ABCA1 Antibody - BSA Free

A: Either DTT or 2-mercaptoethanol is routinely used in SDS-PAGE sample buffer (reducing gel electrophoresis). Except that 2-mercaptoethanol is a stronger reducing agent and is mostly frequently used for that purpose.

A: Please see this link to our protocol for Flow cytometry. This is a general protocol that our lab uses as a guideline.

A: No, this antibody does not have any BSA in its formulation.

A: For some membrane proteins it is not recommended to boil the samples. You may incubate them at room temperature or at 37C in SDS sample buffer with BME. However, from personal experience, if you boil the samples you will not see the protein on the blot. I do not think the exact reason is known.

A: We do not currently have any testing data where we have used NB400-105 with hepatic samples; all of our testing data for NB400-105 can be seen on the product page for this antibody. Should your customer be interested, we do show murine liver data with the following product: NB100-2068.

A: Methanol is not required for antibody binding, but it is a fixation method that we found to not interfere with the antibody-antigen binding. You may use unfixed cells, but you may not be able to achieve staining. If you fix your cells with a formaldehyde-based fixative (we recommend antigen retrieval for IHC). However, as an inter-membrane protein, you will need to permeabilize your cells in order for the antibody to access the protein.

A: The immuogen region of NB400-105 is within residues 1100-1300 of human ABCA1 and as a result it does not cover the extracellular domains, as a consequence, permeabilization is required for this antibody. As a general guideline, we usually recommend using 0.5-1.0% Triton detergent in the buffers in order to allow full permeabilization of our antibody.

A: The post-nuclear lysate you mentioned is posted on our website to demonstrate how we do the experiments, and how it was done by our collaborator (Ref Link #1). The procedure of getting the post-nuclear lysate was just for removing the nuclei before lysis and therefore the cleaner the lysates would be. You do not need to follow the method for the experiment that led to the result.

A: Our own Western blot data for NB400-105 was generated by treating RAW cells with 9-cis-retinoic acid and 22R-hydroxycholesterol, which are both known to induce expression of ABCA1 in macrophages. 40ug of protein was loaded on to the gel in this instance. Unfortunately we do not have any testing data from mouse liver tissues using this particular antibody, however the data for another of our ABCA1 antibodies may be of interest to your customer: NB100-2068. The Western blot data for this product was generated using mouse liver tissue, and a clear band can be seen at the expected molecular weight in wild type tissue. We loaded 30ug of protein when running this Western blot, however, your customer may need to optimise the staining procedure for their sample type.

A: The difference between NB400-105 or NB400-105B is biotin conjugation. It is up to you whether you would like to order the primary antibody that is conjugated or not. With the biotin-conjugated antibody (catalog # NB400-105B), you won't need a secondary, just a (strept)avidin-labelled detection reagent, such as streptavidin-HRP. With the unconjugated primary (catalog # NB400-105), you will need to use a secondary antibody. Some people say that when you use the secondary, there is an amplification of signal, because more than one secondary can bind to each primary. However, with the directly conjugated antibody you have fewer reagents and fewer wash steps.

A: ABCA1 is highly glycosylated and has a predicted molecular weight of approximately 250 kDa without post-translational modification. The apparent molecular weight will change based on treatment and sample. All of the images on our website and datasheet are correct.

A: Most likely the antibody should still work, but we cannot guarantee it. The biggest thing with antibody storage in most cases, especially for unconjugated antibodies, will be to avoid freeze/thaw cycles as much as possible. So even with ones that we recommend to freeze long term we still recommend aliquoting so they can go through as few of those as possible. I would say since it will probably only undergo one when you move it back to 4 degrees Celsius you should be fine, but again I can't say with 100% certainty. If you have a problem with detection using this product, please let us know so that we can figure out other possibilities to help you in that situation.

A: To fractionate a membrane-bound protein, the lipid moieties associated with the protein frequently cause the smearing or streaky appearance of our WB. To result this difficulty, it is advised that we resuspend cell pellets in the SDS-sample buffer, left at room temp without boiling, spinning down the viscous cell debris, and then subject to PAGE and blotting. And this was the reason we suggested you not to boil your protein samples. Whether it will work for resuspending cell pellets in RIPA buffer, unfortunately we have no such data.

A: Either DTT or 2-mercaptoethanol is routinely used in SDS-PAGE sample buffer (reducing gel electrophoresis). Except that 2-mercaptoethanol is a stronger reducing agent and is mostly frequently used for that purpose.

A: Please see this link to our protocol for Flow cytometry. This is a general protocol that our lab uses as a guideline.

A: No, this antibody does not have any BSA in its formulation.

A: For some membrane proteins it is not recommended to boil the samples. You may incubate them at room temperature or at 37C in SDS sample buffer with BME. However, from personal experience, if you boil the samples you will not see the protein on the blot. I do not think the exact reason is known.

A: We do not currently have any testing data where we have used NB400-105 with hepatic samples; all of our testing data for NB400-105 can be seen on the product page for this antibody. Should your customer be interested, we do show murine liver data with the following product: NB100-2068.

A: Methanol is not required for antibody binding, but it is a fixation method that we found to not interfere with the antibody-antigen binding. You may use unfixed cells, but you may not be able to achieve staining. If you fix your cells with a formaldehyde-based fixative (we recommend antigen retrieval for IHC). However, as an inter-membrane protein, you will need to permeabilize your cells in order for the antibody to access the protein.

A: The immuogen region of NB400-105 is within residues 1100-1300 of human ABCA1 and as a result it does not cover the extracellular domains, as a consequence, permeabilization is required for this antibody. As a general guideline, we usually recommend using 0.5-1.0% Triton detergent in the buffers in order to allow full permeabilization of our antibody.

A: The post-nuclear lysate you mentioned is posted on our website to demonstrate how we do the experiments, and how it was done by our collaborator (Ref Link #1). The procedure of getting the post-nuclear lysate was just for removing the nuclei before lysis and therefore the cleaner the lysates would be. You do not need to follow the method for the experiment that led to the result.

A: Our own Western blot data for NB400-105 was generated by treating RAW cells with 9-cis-retinoic acid and 22R-hydroxycholesterol, which are both known to induce expression of ABCA1 in macrophages. 40ug of protein was loaded on to the gel in this instance. Unfortunately we do not have any testing data from mouse liver tissues using this particular antibody, however the data for another of our ABCA1 antibodies may be of interest to your customer: NB100-2068. The Western blot data for this product was generated using mouse liver tissue, and a clear band can be seen at the expected molecular weight in wild type tissue. We loaded 30ug of protein when running this Western blot, however, your customer may need to optimise the staining procedure for their sample type.

A: The difference between NB400-105 or NB400-105B is biotin conjugation. It is up to you whether you would like to order the primary antibody that is conjugated or not. With the biotin-conjugated antibody (catalog # NB400-105B), you won't need a secondary, just a (strept)avidin-labelled detection reagent, such as streptavidin-HRP. With the unconjugated primary (catalog # NB400-105), you will need to use a secondary antibody. Some people say that when you use the secondary, there is an amplification of signal, because more than one secondary can bind to each primary. However, with the directly conjugated antibody you have fewer reagents and fewer wash steps.

A: ABCA1 is highly glycosylated and has a predicted molecular weight of approximately 250 kDa without post-translational modification. The apparent molecular weight will change based on treatment and sample. All of the images on our website and datasheet are correct.

A: Most likely the antibody should still work, but we cannot guarantee it. The biggest thing with antibody storage in most cases, especially for unconjugated antibodies, will be to avoid freeze/thaw cycles as much as possible. So even with ones that we recommend to freeze long term we still recommend aliquoting so they can go through as few of those as possible. I would say since it will probably only undergo one when you move it back to 4 degrees Celsius you should be fine, but again I can't say with 100% certainty. If you have a problem with detection using this product, please let us know so that we can figure out other possibilities to help you in that situation.

A: To fractionate a membrane-bound protein, the lipid moieties associated with the protein frequently cause the smearing or streaky appearance of our WB. To result this difficulty, it is advised that we resuspend cell pellets in the SDS-sample buffer, left at room temp without boiling, spinning down the viscous cell debris, and then subject to PAGE and blotting. And this was the reason we suggested you not to boil your protein samples. Whether it will work for resuspending cell pellets in RIPA buffer, unfortunately we have no such data.

A: Either DTT or 2-mercaptoethanol is routinely used in SDS-PAGE sample buffer (reducing gel electrophoresis). Except that 2-mercaptoethanol is a stronger reducing agent and is mostly frequently used for that purpose.

A: Please see this link to our protocol for Flow cytometry. This is a general protocol that our lab uses as a guideline.

A: No, this antibody does not have any BSA in its formulation.

A: For some membrane proteins it is not recommended to boil the samples. You may incubate them at room temperature or at 37C in SDS sample buffer with BME. However, from personal experience, if you boil the samples you will not see the protein on the blot. I do not think the exact reason is known.

A: We do not currently have any testing data where we have used NB400-105 with hepatic samples; all of our testing data for NB400-105 can be seen on the product page for this antibody. Should your customer be interested, we do show murine liver data with the following product: NB100-2068.

A: Methanol is not required for antibody binding, but it is a fixation method that we found to not interfere with the antibody-antigen binding. You may use unfixed cells, but you may not be able to achieve staining. If you fix your cells with a formaldehyde-based fixative (we recommend antigen retrieval for IHC). However, as an inter-membrane protein, you will need to permeabilize your cells in order for the antibody to access the protein.

A: The immuogen region of NB400-105 is within residues 1100-1300 of human ABCA1 and as a result it does not cover the extracellular domains, as a consequence, permeabilization is required for this antibody. As a general guideline, we usually recommend using 0.5-1.0% Triton detergent in the buffers in order to allow full permeabilization of our antibody.

A: The post-nuclear lysate you mentioned is posted on our website to demonstrate how we do the experiments, and how it was done by our collaborator (Ref Link #1). The procedure of getting the post-nuclear lysate was just for removing the nuclei before lysis and therefore the cleaner the lysates would be. You do not need to follow the method for the experiment that led to the result.

A: Our own Western blot data for NB400-105 was generated by treating RAW cells with 9-cis-retinoic acid and 22R-hydroxycholesterol, which are both known to induce expression of ABCA1 in macrophages. 40ug of protein was loaded on to the gel in this instance. Unfortunately we do not have any testing data from mouse liver tissues using this particular antibody, however the data for another of our ABCA1 antibodies may be of interest to your customer: NB100-2068. The Western blot data for this product was generated using mouse liver tissue, and a clear band can be seen at the expected molecular weight in wild type tissue. We loaded 30ug of protein when running this Western blot, however, your customer may need to optimise the staining procedure for their sample type.

A: The difference between NB400-105 or NB400-105B is biotin conjugation. It is up to you whether you would like to order the primary antibody that is conjugated or not. With the biotin-conjugated antibody (catalog # NB400-105B), you won't need a secondary, just a (strept)avidin-labelled detection reagent, such as streptavidin-HRP. With the unconjugated primary (catalog # NB400-105), you will need to use a secondary antibody. Some people say that when you use the secondary, there is an amplification of signal, because more than one secondary can bind to each primary. However, with the directly conjugated antibody you have fewer reagents and fewer wash steps.

A: ABCA1 is highly glycosylated and has a predicted molecular weight of approximately 250 kDa without post-translational modification. The apparent molecular weight will change based on treatment and sample. All of the images on our website and datasheet are correct.

A: Most likely the antibody should still work, but we cannot guarantee it. The biggest thing with antibody storage in most cases, especially for unconjugated antibodies, will be to avoid freeze/thaw cycles as much as possible. So even with ones that we recommend to freeze long term we still recommend aliquoting so they can go through as few of those as possible. I would say since it will probably only undergo one when you move it back to 4 degrees Celsius you should be fine, but again I can't say with 100% certainty. If you have a problem with detection using this product, please let us know so that we can figure out other possibilities to help you in that situation.

A: To fractionate a membrane-bound protein, the lipid moieties associated with the protein frequently cause the smearing or streaky appearance of our WB. To result this difficulty, it is advised that we resuspend cell pellets in the SDS-sample buffer, left at room temp without boiling, spinning down the viscous cell debris, and then subject to PAGE and blotting. And this was the reason we suggested you not to boil your protein samples. Whether it will work for resuspending cell pellets in RIPA buffer, unfortunately we have no such data.

A: Either DTT or 2-mercaptoethanol is routinely used in SDS-PAGE sample buffer (reducing gel electrophoresis). Except that 2-mercaptoethanol is a stronger reducing agent and is mostly frequently used for that purpose.

A: Please see this link to our protocol for Flow cytometry. This is a general protocol that our lab uses as a guideline.

A: No, this antibody does not have any BSA in its formulation.

A: For some membrane proteins it is not recommended to boil the samples. You may incubate them at room temperature or at 37C in SDS sample buffer with BME. However, from personal experience, if you boil the samples you will not see the protein on the blot. I do not think the exact reason is known.

A: We do not currently have any testing data where we have used NB400-105 with hepatic samples; all of our testing data for NB400-105 can be seen on the product page for this antibody. Should your customer be interested, we do show murine liver data with the following product: NB100-2068.

A: Methanol is not required for antibody binding, but it is a fixation method that we found to not interfere with the antibody-antigen binding. You may use unfixed cells, but you may not be able to achieve staining. If you fix your cells with a formaldehyde-based fixative (we recommend antigen retrieval for IHC). However, as an inter-membrane protein, you will need to permeabilize your cells in order for the antibody to access the protein.

A: The immuogen region of NB400-105 is within residues 1100-1300 of human ABCA1 and as a result it does not cover the extracellular domains, as a consequence, permeabilization is required for this antibody. As a general guideline, we usually recommend using 0.5-1.0% Triton detergent in the buffers in order to allow full permeabilization of our antibody.

A: The post-nuclear lysate you mentioned is posted on our website to demonstrate how we do the experiments, and how it was done by our collaborator (Ref Link #1). The procedure of getting the post-nuclear lysate was just for removing the nuclei before lysis and therefore the cleaner the lysates would be. You do not need to follow the method for the experiment that led to the result.

A: Our own Western blot data for NB400-105 was generated by treating RAW cells with 9-cis-retinoic acid and 22R-hydroxycholesterol, which are both known to induce expression of ABCA1 in macrophages. 40ug of protein was loaded on to the gel in this instance. Unfortunately we do not have any testing data from mouse liver tissues using this particular antibody, however the data for another of our ABCA1 antibodies may be of interest to your customer: NB100-2068. The Western blot data for this product was generated using mouse liver tissue, and a clear band can be seen at the expected molecular weight in wild type tissue. We loaded 30ug of protein when running this Western blot, however, your customer may need to optimise the staining procedure for their sample type.

A: The difference between NB400-105 or NB400-105B is biotin conjugation. It is up to you whether you would like to order the primary antibody that is conjugated or not. With the biotin-conjugated antibody (catalog # NB400-105B), you won't need a secondary, just a (strept)avidin-labelled detection reagent, such as streptavidin-HRP. With the unconjugated primary (catalog # NB400-105), you will need to use a secondary antibody. Some people say that when you use the secondary, there is an amplification of signal, because more than one secondary can bind to each primary. However, with the directly conjugated antibody you have fewer reagents and fewer wash steps.

A: ABCA1 is highly glycosylated and has a predicted molecular weight of approximately 250 kDa without post-translational modification. The apparent molecular weight will change based on treatment and sample. All of the images on our website and datasheet are correct.

A: Most likely the antibody should still work, but we cannot guarantee it. The biggest thing with antibody storage in most cases, especially for unconjugated antibodies, will be to avoid freeze/thaw cycles as much as possible. So even with ones that we recommend to freeze long term we still recommend aliquoting so they can go through as few of those as possible. I would say since it will probably only undergo one when you move it back to 4 degrees Celsius you should be fine, but again I can't say with 100% certainty. If you have a problem with detection using this product, please let us know so that we can figure out other possibilities to help you in that situation.

A: To fractionate a membrane-bound protein, the lipid moieties associated with the protein frequently cause the smearing or streaky appearance of our WB. To result this difficulty, it is advised that we resuspend cell pellets in the SDS-sample buffer, left at room temp without boiling, spinning down the viscous cell debris, and then subject to PAGE and blotting. And this was the reason we suggested you not to boil your protein samples. Whether it will work for resuspending cell pellets in RIPA buffer, unfortunately we have no such data.

A: Either DTT or 2-mercaptoethanol is routinely used in SDS-PAGE sample buffer (reducing gel electrophoresis). Except that 2-mercaptoethanol is a stronger reducing agent and is mostly frequently used for that purpose.

A: Please see this link to our protocol for Flow cytometry. This is a general protocol that our lab uses as a guideline.

A: No, this antibody does not have any BSA in its formulation.

A: For some membrane proteins it is not recommended to boil the samples. You may incubate them at room temperature or at 37C in SDS sample buffer with BME. However, from personal experience, if you boil the samples you will not see the protein on the blot. I do not think the exact reason is known.

A: We do not currently have any testing data where we have used NB400-105 with hepatic samples; all of our testing data for NB400-105 can be seen on the product page for this antibody. Should your customer be interested, we do show murine liver data with the following product: NB100-2068.

A: Methanol is not required for antibody binding, but it is a fixation method that we found to not interfere with the antibody-antigen binding. You may use unfixed cells, but you may not be able to achieve staining. If you fix your cells with a formaldehyde-based fixative (we recommend antigen retrieval for IHC). However, as an inter-membrane protein, you will need to permeabilize your cells in order for the antibody to access the protein.

A: The immuogen region of NB400-105 is within residues 1100-1300 of human ABCA1 and as a result it does not cover the extracellular domains, as a consequence, permeabilization is required for this antibody. As a general guideline, we usually recommend using 0.5-1.0% Triton detergent in the buffers in order to allow full permeabilization of our antibody.

A: The post-nuclear lysate you mentioned is posted on our website to demonstrate how we do the experiments, and how it was done by our collaborator (Ref Link #1). The procedure of getting the post-nuclear lysate was just for removing the nuclei before lysis and therefore the cleaner the lysates would be. You do not need to follow the method for the experiment that led to the result.

A: Our own Western blot data for NB400-105 was generated by treating RAW cells with 9-cis-retinoic acid and 22R-hydroxycholesterol, which are both known to induce expression of ABCA1 in macrophages. 40ug of protein was loaded on to the gel in this instance. Unfortunately we do not have any testing data from mouse liver tissues using this particular antibody, however the data for another of our ABCA1 antibodies may be of interest to your customer: NB100-2068. The Western blot data for this product was generated using mouse liver tissue, and a clear band can be seen at the expected molecular weight in wild type tissue. We loaded 30ug of protein when running this Western blot, however, your customer may need to optimise the staining procedure for their sample type.

A: The difference between NB400-105 or NB400-105B is biotin conjugation. It is up to you whether you would like to order the primary antibody that is conjugated or not. With the biotin-conjugated antibody (catalog # NB400-105B), you won't need a secondary, just a (strept)avidin-labelled detection reagent, such as streptavidin-HRP. With the unconjugated primary (catalog # NB400-105), you will need to use a secondary antibody. Some people say that when you use the secondary, there is an amplification of signal, because more than one secondary can bind to each primary. However, with the directly conjugated antibody you have fewer reagents and fewer wash steps.

A: ABCA1 is highly glycosylated and has a predicted molecular weight of approximately 250 kDa without post-translational modification. The apparent molecular weight will change based on treatment and sample. All of the images on our website and datasheet are correct.

A: Most likely the antibody should still work, but we cannot guarantee it. The biggest thing with antibody storage in most cases, especially for unconjugated antibodies, will be to avoid freeze/thaw cycles as much as possible. So even with ones that we recommend to freeze long term we still recommend aliquoting so they can go through as few of those as possible. I would say since it will probably only undergo one when you move it back to 4 degrees Celsius you should be fine, but again I can't say with 100% certainty. If you have a problem with detection using this product, please let us know so that we can figure out other possibilities to help you in that situation.

A: To fractionate a membrane-bound protein, the lipid moieties associated with the protein frequently cause the smearing or streaky appearance of our WB. To result this difficulty, it is advised that we resuspend cell pellets in the SDS-sample buffer, left at room temp without boiling, spinning down the viscous cell debris, and then subject to PAGE and blotting. And this was the reason we suggested you not to boil your protein samples. Whether it will work for resuspending cell pellets in RIPA buffer, unfortunately we have no such data.

A: Either DTT or 2-mercaptoethanol is routinely used in SDS-PAGE sample buffer (reducing gel electrophoresis). Except that 2-mercaptoethanol is a stronger reducing agent and is mostly frequently used for that purpose.

A: Please see this link to our protocol for Flow cytometry. This is a general protocol that our lab uses as a guideline.

A: No, this antibody does not have any BSA in its formulation.

A: For some membrane proteins it is not recommended to boil the samples. You may incubate them at room temperature or at 37C in SDS sample buffer with BME. However, from personal experience, if you boil the samples you will not see the protein on the blot. I do not think the exact reason is known.

A: We do not currently have any testing data where we have used NB400-105 with hepatic samples; all of our testing data for NB400-105 can be seen on the product page for this antibody. Should your customer be interested, we do show murine liver data with the following product: NB100-2068.

A: Methanol is not required for antibody binding, but it is a fixation method that we found to not interfere with the antibody-antigen binding. You may use unfixed cells, but you may not be able to achieve staining. If you fix your cells with a formaldehyde-based fixative (we recommend antigen retrieval for IHC). However, as an inter-membrane protein, you will need to permeabilize your cells in order for the antibody to access the protein.

A: The immuogen region of NB400-105 is within residues 1100-1300 of human ABCA1 and as a result it does not cover the extracellular domains, as a consequence, permeabilization is required for this antibody. As a general guideline, we usually recommend using 0.5-1.0% Triton detergent in the buffers in order to allow full permeabilization of our antibody.

A: The post-nuclear lysate you mentioned is posted on our website to demonstrate how we do the experiments, and how it was done by our collaborator (Ref Link #1). The procedure of getting the post-nuclear lysate was just for removing the nuclei before lysis and therefore the cleaner the lysates would be. You do not need to follow the method for the experiment that led to the result.

A: Our own Western blot data for NB400-105 was generated by treating RAW cells with 9-cis-retinoic acid and 22R-hydroxycholesterol, which are both known to induce expression of ABCA1 in macrophages. 40ug of protein was loaded on to the gel in this instance. Unfortunately we do not have any testing data from mouse liver tissues using this particular antibody, however the data for another of our ABCA1 antibodies may be of interest to your customer: NB100-2068. The Western blot data for this product was generated using mouse liver tissue, and a clear band can be seen at the expected molecular weight in wild type tissue. We loaded 30ug of protein when running this Western blot, however, your customer may need to optimise the staining procedure for their sample type.

A: The difference between NB400-105 or NB400-105B is biotin conjugation. It is up to you whether you would like to order the primary antibody that is conjugated or not. With the biotin-conjugated antibody (catalog # NB400-105B), you won't need a secondary, just a (strept)avidin-labelled detection reagent, such as streptavidin-HRP. With the unconjugated primary (catalog # NB400-105), you will need to use a secondary antibody. Some people say that when you use the secondary, there is an amplification of signal, because more than one secondary can bind to each primary. However, with the directly conjugated antibody you have fewer reagents and fewer wash steps.

A: ABCA1 is highly glycosylated and has a predicted molecular weight of approximately 250 kDa without post-translational modification. The apparent molecular weight will change based on treatment and sample. All of the images on our website and datasheet are correct.

A: Most likely the antibody should still work, but we cannot guarantee it. The biggest thing with antibody storage in most cases, especially for unconjugated antibodies, will be to avoid freeze/thaw cycles as much as possible. So even with ones that we recommend to freeze long term we still recommend aliquoting so they can go through as few of those as possible. I would say since it will probably only undergo one when you move it back to 4 degrees Celsius you should be fine, but again I can't say with 100% certainty. If you have a problem with detection using this product, please let us know so that we can figure out other possibilities to help you in that situation.

A: To fractionate a membrane-bound protein, the lipid moieties associated with the protein frequently cause the smearing or streaky appearance of our WB. To result this difficulty, it is advised that we resuspend cell pellets in the SDS-sample buffer, left at room temp without boiling, spinning down the viscous cell debris, and then subject to PAGE and blotting. And this was the reason we suggested you not to boil your protein samples. Whether it will work for resuspending cell pellets in RIPA buffer, unfortunately we have no such data.

A: Either DTT or 2-mercaptoethanol is routinely used in SDS-PAGE sample buffer (reducing gel electrophoresis). Except that 2-mercaptoethanol is a stronger reducing agent and is mostly frequently used for that purpose.

A: Please see this link to our protocol for Flow cytometry. This is a general protocol that our lab uses as a guideline.

A: No, this antibody does not have any BSA in its formulation.

A: For some membrane proteins it is not recommended to boil the samples. You may incubate them at room temperature or at 37C in SDS sample buffer with BME. However, from personal experience, if you boil the samples you will not see the protein on the blot. I do not think the exact reason is known.

A: We do not currently have any testing data where we have used NB400-105 with hepatic samples; all of our testing data for NB400-105 can be seen on the product page for this antibody. Should your customer be interested, we do show murine liver data with the following product: NB100-2068.

A: Methanol is not required for antibody binding, but it is a fixation method that we found to not interfere with the antibody-antigen binding. You may use unfixed cells, but you may not be able to achieve staining. If you fix your cells with a formaldehyde-based fixative (we recommend antigen retrieval for IHC). However, as an inter-membrane protein, you will need to permeabilize your cells in order for the antibody to access the protein.

A: The immuogen region of NB400-105 is within residues 1100-1300 of human ABCA1 and as a result it does not cover the extracellular domains, as a consequence, permeabilization is required for this antibody. As a general guideline, we usually recommend using 0.5-1.0% Triton detergent in the buffers in order to allow full permeabilization of our antibody.

A: The post-nuclear lysate you mentioned is posted on our website to demonstrate how we do the experiments, and how it was done by our collaborator (Ref Link #1). The procedure of getting the post-nuclear lysate was just for removing the nuclei before lysis and therefore the cleaner the lysates would be. You do not need to follow the method for the experiment that led to the result.

A: Our own Western blot data for NB400-105 was generated by treating RAW cells with 9-cis-retinoic acid and 22R-hydroxycholesterol, which are both known to induce expression of ABCA1 in macrophages. 40ug of protein was loaded on to the gel in this instance. Unfortunately we do not have any testing data from mouse liver tissues using this particular antibody, however the data for another of our ABCA1 antibodies may be of interest to your customer: NB100-2068. The Western blot data for this product was generated using mouse liver tissue, and a clear band can be seen at the expected molecular weight in wild type tissue. We loaded 30ug of protein when running this Western blot, however, your customer may need to optimise the staining procedure for their sample type.

A: The difference between NB400-105 or NB400-105B is biotin conjugation. It is up to you whether you would like to order the primary antibody that is conjugated or not. With the biotin-conjugated antibody (catalog # NB400-105B), you won't need a secondary, just a (strept)avidin-labelled detection reagent, such as streptavidin-HRP. With the unconjugated primary (catalog # NB400-105), you will need to use a secondary antibody. Some people say that when you use the secondary, there is an amplification of signal, because more than one secondary can bind to each primary. However, with the directly conjugated antibody you have fewer reagents and fewer wash steps.

A: ABCA1 is highly glycosylated and has a predicted molecular weight of approximately 250 kDa without post-translational modification. The apparent molecular weight will change based on treatment and sample. All of the images on our website and datasheet are correct.

A: Most likely the antibody should still work, but we cannot guarantee it. The biggest thing with antibody storage in most cases, especially for unconjugated antibodies, will be to avoid freeze/thaw cycles as much as possible. So even with ones that we recommend to freeze long term we still recommend aliquoting so they can go through as few of those as possible. I would say since it will probably only undergo one when you move it back to 4 degrees Celsius you should be fine, but again I can't say with 100% certainty. If you have a problem with detection using this product, please let us know so that we can figure out other possibilities to help you in that situation.

A: To fractionate a membrane-bound protein, the lipid moieties associated with the protein frequently cause the smearing or streaky appearance of our WB. To result this difficulty, it is advised that we resuspend cell pellets in the SDS-sample buffer, left at room temp without boiling, spinning down the viscous cell debris, and then subject to PAGE and blotting. And this was the reason we suggested you not to boil your protein samples. Whether it will work for resuspending cell pellets in RIPA buffer, unfortunately we have no such data.

A: Either DTT or 2-mercaptoethanol is routinely used in SDS-PAGE sample buffer (reducing gel electrophoresis). Except that 2-mercaptoethanol is a stronger reducing agent and is mostly frequently used for that purpose.

A: Please see this link to our protocol for Flow cytometry. This is a general protocol that our lab uses as a guideline.

A: No, this antibody does not have any BSA in its formulation.

A: For some membrane proteins it is not recommended to boil the samples. You may incubate them at room temperature or at 37C in SDS sample buffer with BME. However, from personal experience, if you boil the samples you will not see the protein on the blot. I do not think the exact reason is known.

A: We do not currently have any testing data where we have used NB400-105 with hepatic samples; all of our testing data for NB400-105 can be seen on the product page for this antibody. Should your customer be interested, we do show murine liver data with the following product: NB100-2068.

A: Methanol is not required for antibody binding, but it is a fixation method that we found to not interfere with the antibody-antigen binding. You may use unfixed cells, but you may not be able to achieve staining. If you fix your cells with a formaldehyde-based fixative (we recommend antigen retrieval for IHC). However, as an inter-membrane protein, you will need to permeabilize your cells in order for the antibody to access the protein.

A: The immuogen region of NB400-105 is within residues 1100-1300 of human ABCA1 and as a result it does not cover the extracellular domains, as a consequence, permeabilization is required for this antibody. As a general guideline, we usually recommend using 0.5-1.0% Triton detergent in the buffers in order to allow full permeabilization of our antibody.

A: The post-nuclear lysate you mentioned is posted on our website to demonstrate how we do the experiments, and how it was done by our collaborator (Ref Link #1). The procedure of getting the post-nuclear lysate was just for removing the nuclei before lysis and therefore the cleaner the lysates would be. You do not need to follow the method for the experiment that led to the result.

A: Our own Western blot data for NB400-105 was generated by treating RAW cells with 9-cis-retinoic acid and 22R-hydroxycholesterol, which are both known to induce expression of ABCA1 in macrophages. 40ug of protein was loaded on to the gel in this instance. Unfortunately we do not have any testing data from mouse liver tissues using this particular antibody, however the data for another of our ABCA1 antibodies may be of interest to your customer: NB100-2068. The Western blot data for this product was generated using mouse liver tissue, and a clear band can be seen at the expected molecular weight in wild type tissue. We loaded 30ug of protein when running this Western blot, however, your customer may need to optimise the staining procedure for their sample type.

A: The difference between NB400-105 or NB400-105B is biotin conjugation. It is up to you whether you would like to order the primary antibody that is conjugated or not. With the biotin-conjugated antibody (catalog # NB400-105B), you won't need a secondary, just a (strept)avidin-labelled detection reagent, such as streptavidin-HRP. With the unconjugated primary (catalog # NB400-105), you will need to use a secondary antibody. Some people say that when you use the secondary, there is an amplification of signal, because more than one secondary can bind to each primary. However, with the directly conjugated antibody you have fewer reagents and fewer wash steps.

A: ABCA1 is highly glycosylated and has a predicted molecular weight of approximately 250 kDa without post-translational modification. The apparent molecular weight will change based on treatment and sample. All of the images on our website and datasheet are correct.

A: Most likely the antibody should still work, but we cannot guarantee it. The biggest thing with antibody storage in most cases, especially for unconjugated antibodies, will be to avoid freeze/thaw cycles as much as possible. So even with ones that we recommend to freeze long term we still recommend aliquoting so they can go through as few of those as possible. I would say since it will probably only undergo one when you move it back to 4 degrees Celsius you should be fine, but again I can't say with 100% certainty. If you have a problem with detection using this product, please let us know so that we can figure out other possibilities to help you in that situation.

A: To fractionate a membrane-bound protein, the lipid moieties associated with the protein frequently cause the smearing or streaky appearance of our WB. To result this difficulty, it is advised that we resuspend cell pellets in the SDS-sample buffer, left at room temp without boiling, spinning down the viscous cell debris, and then subject to PAGE and blotting. And this was the reason we suggested you not to boil your protein samples. Whether it will work for resuspending cell pellets in RIPA buffer, unfortunately we have no such data.

A: Either DTT or 2-mercaptoethanol is routinely used in SDS-PAGE sample buffer (reducing gel electrophoresis). Except that 2-mercaptoethanol is a stronger reducing agent and is mostly frequently used for that purpose.

A: Please see this link to our protocol for Flow cytometry. This is a general protocol that our lab uses as a guideline.

A: No, this antibody does not have any BSA in its formulation.

A: For some membrane proteins it is not recommended to boil the samples. You may incubate them at room temperature or at 37C in SDS sample buffer with BME. However, from personal experience, if you boil the samples you will not see the protein on the blot. I do not think the exact reason is known.

A: We do not currently have any testing data where we have used NB400-105 with hepatic samples; all of our testing data for NB400-105 can be seen on the product page for this antibody. Should your customer be interested, we do show murine liver data with the following product: NB100-2068.

A: Methanol is not required for antibody binding, but it is a fixation method that we found to not interfere with the antibody-antigen binding. You may use unfixed cells, but you may not be able to achieve staining. If you fix your cells with a formaldehyde-based fixative (we recommend antigen retrieval for IHC). However, as an inter-membrane protein, you will need to permeabilize your cells in order for the antibody to access the protein.

A: The immuogen region of NB400-105 is within residues 1100-1300 of human ABCA1 and as a result it does not cover the extracellular domains, as a consequence, permeabilization is required for this antibody. As a general guideline, we usually recommend using 0.5-1.0% Triton detergent in the buffers in order to allow full permeabilization of our antibody.

A: The post-nuclear lysate you mentioned is posted on our website to demonstrate how we do the experiments, and how it was done by our collaborator (Ref Link #1). The procedure of getting the post-nuclear lysate was just for removing the nuclei before lysis and therefore the cleaner the lysates would be. You do not need to follow the method for the experiment that led to the result.

A: Our own Western blot data for NB400-105 was generated by treating RAW cells with 9-cis-retinoic acid and 22R-hydroxycholesterol, which are both known to induce expression of ABCA1 in macrophages. 40ug of protein was loaded on to the gel in this instance. Unfortunately we do not have any testing data from mouse liver tissues using this particular antibody, however the data for another of our ABCA1 antibodies may be of interest to your customer: NB100-2068. The Western blot data for this product was generated using mouse liver tissue, and a clear band can be seen at the expected molecular weight in wild type tissue. We loaded 30ug of protein when running this Western blot, however, your customer may need to optimise the staining procedure for their sample type.

A: The difference between NB400-105 or NB400-105B is biotin conjugation. It is up to you whether you would like to order the primary antibody that is conjugated or not. With the biotin-conjugated antibody (catalog # NB400-105B), you won't need a secondary, just a (strept)avidin-labelled detection reagent, such as streptavidin-HRP. With the unconjugated primary (catalog # NB400-105), you will need to use a secondary antibody. Some people say that when you use the secondary, there is an amplification of signal, because more than one secondary can bind to each primary. However, with the directly conjugated antibody you have fewer reagents and fewer wash steps.

A: ABCA1 is highly glycosylated and has a predicted molecular weight of approximately 250 kDa without post-translational modification. The apparent molecular weight will change based on treatment and sample. All of the images on our website and datasheet are correct.

A: Most likely the antibody should still work, but we cannot guarantee it. The biggest thing with antibody storage in most cases, especially for unconjugated antibodies, will be to avoid freeze/thaw cycles as much as possible. So even with ones that we recommend to freeze long term we still recommend aliquoting so they can go through as few of those as possible. I would say since it will probably only undergo one when you move it back to 4 degrees Celsius you should be fine, but again I can't say with 100% certainty. If you have a problem with detection using this product, please let us know so that we can figure out other possibilities to help you in that situation.

A: To fractionate a membrane-bound protein, the lipid moieties associated with the protein frequently cause the smearing or streaky appearance of our WB. To result this difficulty, it is advised that we resuspend cell pellets in the SDS-sample buffer, left at room temp without boiling, spinning down the viscous cell debris, and then subject to PAGE and blotting. And this was the reason we suggested you not to boil your protein samples. Whether it will work for resuspending cell pellets in RIPA buffer, unfortunately we have no such data.

A: Either DTT or 2-mercaptoethanol is routinely used in SDS-PAGE sample buffer (reducing gel electrophoresis). Except that 2-mercaptoethanol is a stronger reducing agent and is mostly frequently used for that purpose.

A: Please see this link to our protocol for Flow cytometry. This is a general protocol that our lab uses as a guideline.

A: No, this antibody does not have any BSA in its formulation.

A: For some membrane proteins it is not recommended to boil the samples. You may incubate them at room temperature or at 37C in SDS sample buffer with BME. However, from personal experience, if you boil the samples you will not see the protein on the blot. I do not think the exact reason is known.

A: We do not currently have any testing data where we have used NB400-105 with hepatic samples; all of our testing data for NB400-105 can be seen on the product page for this antibody. Should your customer be interested, we do show murine liver data with the following product: NB100-2068.

A: Methanol is not required for antibody binding, but it is a fixation method that we found to not interfere with the antibody-antigen binding. You may use unfixed cells, but you may not be able to achieve staining. If you fix your cells with a formaldehyde-based fixative (we recommend antigen retrieval for IHC). However, as an inter-membrane protein, you will need to permeabilize your cells in order for the antibody to access the protein.

A: The immuogen region of NB400-105 is within residues 1100-1300 of human ABCA1 and as a result it does not cover the extracellular domains, as a consequence, permeabilization is required for this antibody. As a general guideline, we usually recommend using 0.5-1.0% Triton detergent in the buffers in order to allow full permeabilization of our antibody.

A: The post-nuclear lysate you mentioned is posted on our website to demonstrate how we do the experiments, and how it was done by our collaborator (Ref Link #1). The procedure of getting the post-nuclear lysate was just for removing the nuclei before lysis and therefore the cleaner the lysates would be. You do not need to follow the method for the experiment that led to the result.

A: Our own Western blot data for NB400-105 was generated by treating RAW cells with 9-cis-retinoic acid and 22R-hydroxycholesterol, which are both known to induce expression of ABCA1 in macrophages. 40ug of protein was loaded on to the gel in this instance. Unfortunately we do not have any testing data from mouse liver tissues using this particular antibody, however the data for another of our ABCA1 antibodies may be of interest to your customer: NB100-2068. The Western blot data for this product was generated using mouse liver tissue, and a clear band can be seen at the expected molecular weight in wild type tissue. We loaded 30ug of protein when running this Western blot, however, your customer may need to optimise the staining procedure for their sample type.

A: The difference between NB400-105 or NB400-105B is biotin conjugation. It is up to you whether you would like to order the primary antibody that is conjugated or not. With the biotin-conjugated antibody (catalog # NB400-105B), you won't need a secondary, just a (strept)avidin-labelled detection reagent, such as streptavidin-HRP. With the unconjugated primary (catalog # NB400-105), you will need to use a secondary antibody. Some people say that when you use the secondary, there is an amplification of signal, because more than one secondary can bind to each primary. However, with the directly conjugated antibody you have fewer reagents and fewer wash steps.

A: ABCA1 is highly glycosylated and has a predicted molecular weight of approximately 250 kDa without post-translational modification. The apparent molecular weight will change based on treatment and sample. All of the images on our website and datasheet are correct.

A: Most likely the antibody should still work, but we cannot guarantee it. The biggest thing with antibody storage in most cases, especially for unconjugated antibodies, will be to avoid freeze/thaw cycles as much as possible. So even with ones that we recommend to freeze long term we still recommend aliquoting so they can go through as few of those as possible. I would say since it will probably only undergo one when you move it back to 4 degrees Celsius you should be fine, but again I can't say with 100% certainty. If you have a problem with detection using this product, please let us know so that we can figure out other possibilities to help you in that situation.

A: To fractionate a membrane-bound protein, the lipid moieties associated with the protein frequently cause the smearing or streaky appearance of our WB. To result this difficulty, it is advised that we resuspend cell pellets in the SDS-sample buffer, left at room temp without boiling, spinning down the viscous cell debris, and then subject to PAGE and blotting. And this was the reason we suggested you not to boil your protein samples. Whether it will work for resuspending cell pellets in RIPA buffer, unfortunately we have no such data.

A: Either DTT or 2-mercaptoethanol is routinely used in SDS-PAGE sample buffer (reducing gel electrophoresis). Except that 2-mercaptoethanol is a stronger reducing agent and is mostly frequently used for that purpose.

A: Please see this link to our protocol for Flow cytometry. This is a general protocol that our lab uses as a guideline.

A: No, this antibody does not have any BSA in its formulation.

A: For some membrane proteins it is not recommended to boil the samples. You may incubate them at room temperature or at 37C in SDS sample buffer with BME. However, from personal experience, if you boil the samples you will not see the protein on the blot. I do not think the exact reason is known.

A: We do not currently have any testing data where we have used NB400-105 with hepatic samples; all of our testing data for NB400-105 can be seen on the product page for this antibody. Should your customer be interested, we do show murine liver data with the following product: NB100-2068.

A: Methanol is not required for antibody binding, but it is a fixation method that we found to not interfere with the antibody-antigen binding. You may use unfixed cells, but you may not be able to achieve staining. If you fix your cells with a formaldehyde-based fixative (we recommend antigen retrieval for IHC). However, as an inter-membrane protein, you will need to permeabilize your cells in order for the antibody to access the protein.

A: The immuogen region of NB400-105 is within residues 1100-1300 of human ABCA1 and as a result it does not cover the extracellular domains, as a consequence, permeabilization is required for this antibody. As a general guideline, we usually recommend using 0.5-1.0% Triton detergent in the buffers in order to allow full permeabilization of our antibody.

A: The post-nuclear lysate you mentioned is posted on our website to demonstrate how we do the experiments, and how it was done by our collaborator (Ref Link #1). The procedure of getting the post-nuclear lysate was just for removing the nuclei before lysis and therefore the cleaner the lysates would be. You do not need to follow the method for the experiment that led to the result.

A: Our own Western blot data for NB400-105 was generated by treating RAW cells with 9-cis-retinoic acid and 22R-hydroxycholesterol, which are both known to induce expression of ABCA1 in macrophages. 40ug of protein was loaded on to the gel in this instance. Unfortunately we do not have any testing data from mouse liver tissues using this particular antibody, however the data for another of our ABCA1 antibodies may be of interest to your customer: NB100-2068. The Western blot data for this product was generated using mouse liver tissue, and a clear band can be seen at the expected molecular weight in wild type tissue. We loaded 30ug of protein when running this Western blot, however, your customer may need to optimise the staining procedure for their sample type.

A: The difference between NB400-105 or NB400-105B is biotin conjugation. It is up to you whether you would like to order the primary antibody that is conjugated or not. With the biotin-conjugated antibody (catalog # NB400-105B), you won't need a secondary, just a (strept)avidin-labelled detection reagent, such as streptavidin-HRP. With the unconjugated primary (catalog # NB400-105), you will need to use a secondary antibody. Some people say that when you use the secondary, there is an amplification of signal, because more than one secondary can bind to each primary. However, with the directly conjugated antibody you have fewer reagents and fewer wash steps.

A: ABCA1 is highly glycosylated and has a predicted molecular weight of approximately 250 kDa without post-translational modification. The apparent molecular weight will change based on treatment and sample. All of the images on our website and datasheet are correct.

A: Most likely the antibody should still work, but we cannot guarantee it. The biggest thing with antibody storage in most cases, especially for unconjugated antibodies, will be to avoid freeze/thaw cycles as much as possible. So even with ones that we recommend to freeze long term we still recommend aliquoting so they can go through as few of those as possible. I would say since it will probably only undergo one when you move it back to 4 degrees Celsius you should be fine, but again I can't say with 100% certainty. If you have a problem with detection using this product, please let us know so that we can figure out other possibilities to help you in that situation.

A: To fractionate a membrane-bound protein, the lipid moieties associated with the protein frequently cause the smearing or streaky appearance of our WB. To result this difficulty, it is advised that we resuspend cell pellets in the SDS-sample buffer, left at room temp without boiling, spinning down the viscous cell debris, and then subject to PAGE and blotting. And this was the reason we suggested you not to boil your protein samples. Whether it will work for resuspending cell pellets in RIPA buffer, unfortunately we have no such data.

A: Either DTT or 2-mercaptoethanol is routinely used in SDS-PAGE sample buffer (reducing gel electrophoresis). Except that 2-mercaptoethanol is a stronger reducing agent and is mostly frequently used for that purpose.

A: Please see this link to our protocol for Flow cytometry. This is a general protocol that our lab uses as a guideline.

A: No, this antibody does not have any BSA in its formulation.

A: For some membrane proteins it is not recommended to boil the samples. You may incubate them at room temperature or at 37C in SDS sample buffer with BME. However, from personal experience, if you boil the samples you will not see the protein on the blot. I do not think the exact reason is known.

A: We do not currently have any testing data where we have used NB400-105 with hepatic samples; all of our testing data for NB400-105 can be seen on the product page for this antibody. Should your customer be interested, we do show murine liver data with the following product: NB100-2068.

A: Methanol is not required for antibody binding, but it is a fixation method that we found to not interfere with the antibody-antigen binding. You may use unfixed cells, but you may not be able to achieve staining. If you fix your cells with a formaldehyde-based fixative (we recommend antigen retrieval for IHC). However, as an inter-membrane protein, you will need to permeabilize your cells in order for the antibody to access the protein.

A: The immuogen region of NB400-105 is within residues 1100-1300 of human ABCA1 and as a result it does not cover the extracellular domains, as a consequence, permeabilization is required for this antibody. As a general guideline, we usually recommend using 0.5-1.0% Triton detergent in the buffers in order to allow full permeabilization of our antibody.

A: The post-nuclear lysate you mentioned is posted on our website to demonstrate how we do the experiments, and how it was done by our collaborator (Ref Link #1). The procedure of getting the post-nuclear lysate was just for removing the nuclei before lysis and therefore the cleaner the lysates would be. You do not need to follow the method for the experiment that led to the result.

A: Our own Western blot data for NB400-105 was generated by treating RAW cells with 9-cis-retinoic acid and 22R-hydroxycholesterol, which are both known to induce expression of ABCA1 in macrophages. 40ug of protein was loaded on to the gel in this instance. Unfortunately we do not have any testing data from mouse liver tissues using this particular antibody, however the data for another of our ABCA1 antibodies may be of interest to your customer: NB100-2068. The Western blot data for this product was generated using mouse liver tissue, and a clear band can be seen at the expected molecular weight in wild type tissue. We loaded 30ug of protein when running this Western blot, however, your customer may need to optimise the staining procedure for their sample type.

A: The difference between NB400-105 or NB400-105B is biotin conjugation. It is up to you whether you would like to order the primary antibody that is conjugated or not. With the biotin-conjugated antibody (catalog # NB400-105B), you won't need a secondary, just a (strept)avidin-labelled detection reagent, such as streptavidin-HRP. With the unconjugated primary (catalog # NB400-105), you will need to use a secondary antibody. Some people say that when you use the secondary, there is an amplification of signal, because more than one secondary can bind to each primary. However, with the directly conjugated antibody you have fewer reagents and fewer wash steps.

A: ABCA1 is highly glycosylated and has a predicted molecular weight of approximately 250 kDa without post-translational modification. The apparent molecular weight will change based on treatment and sample. All of the images on our website and datasheet are correct.

A: Most likely the antibody should still work, but we cannot guarantee it. The biggest thing with antibody storage in most cases, especially for unconjugated antibodies, will be to avoid freeze/thaw cycles as much as possible. So even with ones that we recommend to freeze long term we still recommend aliquoting so they can go through as few of those as possible. I would say since it will probably only undergo one when you move it back to 4 degrees Celsius you should be fine, but again I can't say with 100% certainty. If you have a problem with detection using this product, please let us know so that we can figure out other possibilities to help you in that situation.

A: To fractionate a membrane-bound protein, the lipid moieties associated with the protein frequently cause the smearing or streaky appearance of our WB. To result this difficulty, it is advised that we resuspend cell pellets in the SDS-sample buffer, left at room temp without boiling, spinning down the viscous cell debris, and then subject to PAGE and blotting. And this was the reason we suggested you not to boil your protein samples. Whether it will work for resuspending cell pellets in RIPA buffer, unfortunately we have no such data.

A: Either DTT or 2-mercaptoethanol is routinely used in SDS-PAGE sample buffer (reducing gel electrophoresis). Except that 2-mercaptoethanol is a stronger reducing agent and is mostly frequently used for that purpose.

A: Please see this link to our protocol for Flow cytometry. This is a general protocol that our lab uses as a guideline.

A: No, this antibody does not have any BSA in its formulation.

A: For some membrane proteins it is not recommended to boil the samples. You may incubate them at room temperature or at 37C in SDS sample buffer with BME. However, from personal experience, if you boil the samples you will not see the protein on the blot. I do not think the exact reason is known.

A: We do not currently have any testing data where we have used NB400-105 with hepatic samples; all of our testing data for NB400-105 can be seen on the product page for this antibody. Should your customer be interested, we do show murine liver data with the following product: NB100-2068.

A: Methanol is not required for antibody binding, but it is a fixation method that we found to not interfere with the antibody-antigen binding. You may use unfixed cells, but you may not be able to achieve staining. If you fix your cells with a formaldehyde-based fixative (we recommend antigen retrieval for IHC). However, as an inter-membrane protein, you will need to permeabilize your cells in order for the antibody to access the protein.

A: The immuogen region of NB400-105 is within residues 1100-1300 of human ABCA1 and as a result it does not cover the extracellular domains, as a consequence, permeabilization is required for this antibody. As a general guideline, we usually recommend using 0.5-1.0% Triton detergent in the buffers in order to allow full permeabilization of our antibody.

A: The post-nuclear lysate you mentioned is posted on our website to demonstrate how we do the experiments, and how it was done by our collaborator (Ref Link #1). The procedure of getting the post-nuclear lysate was just for removing the nuclei before lysis and therefore the cleaner the lysates would be. You do not need to follow the method for the experiment that led to the result.

A: Our own Western blot data for NB400-105 was generated by treating RAW cells with 9-cis-retinoic acid and 22R-hydroxycholesterol, which are both known to induce expression of ABCA1 in macrophages. 40ug of protein was loaded on to the gel in this instance. Unfortunately we do not have any testing data from mouse liver tissues using this particular antibody, however the data for another of our ABCA1 antibodies may be of interest to your customer: NB100-2068. The Western blot data for this product was generated using mouse liver tissue, and a clear band can be seen at the expected molecular weight in wild type tissue. We loaded 30ug of protein when running this Western blot, however, your customer may need to optimise the staining procedure for their sample type.

A: The difference between NB400-105 or NB400-105B is biotin conjugation. It is up to you whether you would like to order the primary antibody that is conjugated or not. With the biotin-conjugated antibody (catalog # NB400-105B), you won't need a secondary, just a (strept)avidin-labelled detection reagent, such as streptavidin-HRP. With the unconjugated primary (catalog # NB400-105), you will need to use a secondary antibody. Some people say that when you use the secondary, there is an amplification of signal, because more than one secondary can bind to each primary. However, with the directly conjugated antibody you have fewer reagents and fewer wash steps.

A: ABCA1 is highly glycosylated and has a predicted molecular weight of approximately 250 kDa without post-translational modification. The apparent molecular weight will change based on treatment and sample. All of the images on our website and datasheet are correct.

A: Most likely the antibody should still work, but we cannot guarantee it. The biggest thing with antibody storage in most cases, especially for unconjugated antibodies, will be to avoid freeze/thaw cycles as much as possible. So even with ones that we recommend to freeze long term we still recommend aliquoting so they can go through as few of those as possible. I would say since it will probably only undergo one when you move it back to 4 degrees Celsius you should be fine, but again I can't say with 100% certainty. If you have a problem with detection using this product, please let us know so that we can figure out other possibilities to help you in that situation.

A: To fractionate a membrane-bound protein, the lipid moieties associated with the protein frequently cause the smearing or streaky appearance of our WB. To result this difficulty, it is advised that we resuspend cell pellets in the SDS-sample buffer, left at room temp without boiling, spinning down the viscous cell debris, and then subject to PAGE and blotting. And this was the reason we suggested you not to boil your protein samples. Whether it will work for resuspending cell pellets in RIPA buffer, unfortunately we have no such data.