Endoplasmic Reticulum Enrichment Extraction Kit

A: The kit components for our kit NBP2-29482 do not contain any ascorbic acid. The only component that contains any acids is KC-121, and this component contains 1% acetic acid.

A: The short answer is Yes, this kit NBP2-29482 (Endoplasmic Reticulum Enrichment Extraction Kit) can be used in all cell lines. The yielding information/rate can also be obtained on our web datasheet. Endoplasmic Reticulum Enrichment Extraction Kit (NBP2-29482) datasheet, under the Title of "Applications/Dilutions".

A:

Our kit NBP2-29482 does not contain any polyclonal antibodies. The components list for this kit can be found on page five of the manual

A: We recommend you to review paper entitled "A rapid calcium precipitation method of recovering large amounts of highly pure hepatocyte rough endoplasmic reticulum" by Hamilton et al ( published in Journal of Lipid Research Volume 40, 1999) wherein the modified calcium precipitation method had been described in more detail. The 5x CaCl2 solution is 40mM. You may be able to improve your results by using ice-cold 1x (8mM) CaCl2 and adding it drop-wise to the solution. The GRP78 protein is a ER marker but it will mark both smooth and rough ER, so the fact that there is plenty of GRP78 in your solution does not have to be an indication that the extraction was not complete. There are just smooth ER vesicles in the solution, which won't get precipitated until you spin it down at min 90 000xg.

A: With regard to your inquiry, my lab thinks that if the lysosomes are the same density and size as the ER population, they will be present in the ER collection. They recommend additional homogenization/lysis time on your cell population at the beginning of your enrichment process. They also think that further dilution could help as well. If possible, also try to add another centrifuge step to help remove other organelles prior to the high speed spin. We think these suggestions should help to eliminate the presence of the lysosomes in your ER collection.

A: We have evaluated this Kit in tissues. We have not yet tested this Kit in cell lines. However, this kit has been described for use with cell lines in citation #2 in the product manual.

A: We use the Wheaton grinder loose version. With our centrifuge we set the speed at 90000 g, our rotor doesn't have options to specify max or average settings, there is just a setting. But if the options on your rotor are for max or average, then I would imagine average would be the one to choose, but either one might work just as well.

A: We have used the kit with tissue. We haven't used the kit with cells. The customer will need to empirically determine the amount of cells needed. We unfortunately do not have any information on whether or not detergents will affect the usefulness of this kit. This kit is designed for endoplasmic reticulum enrichment. It is not designed for preservation of the Golgi and cell membrane fractions. The kit has been developed based on information in the general references listed in the manual provided with the kit.

A: The kit components for our kit NBP2-29482 do not contain any ascorbic acid. The only component that contains any acids is KC-121, and this component contains 1% acetic acid.

A: The short answer is Yes, this kit NBP2-29482 (Endoplasmic Reticulum Enrichment Extraction Kit) can be used in all cell lines. The yielding information/rate can also be obtained on our web datasheet. Endoplasmic Reticulum Enrichment Extraction Kit (NBP2-29482) datasheet, under the Title of "Applications/Dilutions".

A:

Our kit NBP2-29482 does not contain any polyclonal antibodies. The components list for this kit can be found on page five of the manual

A: We recommend you to review paper entitled "A rapid calcium precipitation method of recovering large amounts of highly pure hepatocyte rough endoplasmic reticulum" by Hamilton et al ( published in Journal of Lipid Research Volume 40, 1999) wherein the modified calcium precipitation method had been described in more detail. The 5x CaCl2 solution is 40mM. You may be able to improve your results by using ice-cold 1x (8mM) CaCl2 and adding it drop-wise to the solution. The GRP78 protein is a ER marker but it will mark both smooth and rough ER, so the fact that there is plenty of GRP78 in your solution does not have to be an indication that the extraction was not complete. There are just smooth ER vesicles in the solution, which won't get precipitated until you spin it down at min 90 000xg.

A: With regard to your inquiry, my lab thinks that if the lysosomes are the same density and size as the ER population, they will be present in the ER collection. They recommend additional homogenization/lysis time on your cell population at the beginning of your enrichment process. They also think that further dilution could help as well. If possible, also try to add another centrifuge step to help remove other organelles prior to the high speed spin. We think these suggestions should help to eliminate the presence of the lysosomes in your ER collection.

A: We have evaluated this Kit in tissues. We have not yet tested this Kit in cell lines. However, this kit has been described for use with cell lines in citation #2 in the product manual.

A: We use the Wheaton grinder loose version. With our centrifuge we set the speed at 90000 g, our rotor doesn't have options to specify max or average settings, there is just a setting. But if the options on your rotor are for max or average, then I would imagine average would be the one to choose, but either one might work just as well.

A: We have used the kit with tissue. We haven't used the kit with cells. The customer will need to empirically determine the amount of cells needed. We unfortunately do not have any information on whether or not detergents will affect the usefulness of this kit. This kit is designed for endoplasmic reticulum enrichment. It is not designed for preservation of the Golgi and cell membrane fractions. The kit has been developed based on information in the general references listed in the manual provided with the kit.

A: The kit components for our kit NBP2-29482 do not contain any ascorbic acid. The only component that contains any acids is KC-121, and this component contains 1% acetic acid.

A: The short answer is Yes, this kit NBP2-29482 (Endoplasmic Reticulum Enrichment Extraction Kit) can be used in all cell lines. The yielding information/rate can also be obtained on our web datasheet. Endoplasmic Reticulum Enrichment Extraction Kit (NBP2-29482) datasheet, under the Title of "Applications/Dilutions".

A:

Our kit NBP2-29482 does not contain any polyclonal antibodies. The components list for this kit can be found on page five of the manual

A: We recommend you to review paper entitled "A rapid calcium precipitation method of recovering large amounts of highly pure hepatocyte rough endoplasmic reticulum" by Hamilton et al ( published in Journal of Lipid Research Volume 40, 1999) wherein the modified calcium precipitation method had been described in more detail. The 5x CaCl2 solution is 40mM. You may be able to improve your results by using ice-cold 1x (8mM) CaCl2 and adding it drop-wise to the solution. The GRP78 protein is a ER marker but it will mark both smooth and rough ER, so the fact that there is plenty of GRP78 in your solution does not have to be an indication that the extraction was not complete. There are just smooth ER vesicles in the solution, which won't get precipitated until you spin it down at min 90 000xg.

A: With regard to your inquiry, my lab thinks that if the lysosomes are the same density and size as the ER population, they will be present in the ER collection. They recommend additional homogenization/lysis time on your cell population at the beginning of your enrichment process. They also think that further dilution could help as well. If possible, also try to add another centrifuge step to help remove other organelles prior to the high speed spin. We think these suggestions should help to eliminate the presence of the lysosomes in your ER collection.

A: We have evaluated this Kit in tissues. We have not yet tested this Kit in cell lines. However, this kit has been described for use with cell lines in citation #2 in the product manual.

A: We use the Wheaton grinder loose version. With our centrifuge we set the speed at 90000 g, our rotor doesn't have options to specify max or average settings, there is just a setting. But if the options on your rotor are for max or average, then I would imagine average would be the one to choose, but either one might work just as well.

A: We have used the kit with tissue. We haven't used the kit with cells. The customer will need to empirically determine the amount of cells needed. We unfortunately do not have any information on whether or not detergents will affect the usefulness of this kit. This kit is designed for endoplasmic reticulum enrichment. It is not designed for preservation of the Golgi and cell membrane fractions. The kit has been developed based on information in the general references listed in the manual provided with the kit.

A: The kit components for our kit NBP2-29482 do not contain any ascorbic acid. The only component that contains any acids is KC-121, and this component contains 1% acetic acid.

A: The short answer is Yes, this kit NBP2-29482 (Endoplasmic Reticulum Enrichment Extraction Kit) can be used in all cell lines. The yielding information/rate can also be obtained on our web datasheet. Endoplasmic Reticulum Enrichment Extraction Kit (NBP2-29482) datasheet, under the Title of "Applications/Dilutions".

A:

Our kit NBP2-29482 does not contain any polyclonal antibodies. The components list for this kit can be found on page five of the manual

A: We recommend you to review paper entitled "A rapid calcium precipitation method of recovering large amounts of highly pure hepatocyte rough endoplasmic reticulum" by Hamilton et al ( published in Journal of Lipid Research Volume 40, 1999) wherein the modified calcium precipitation method had been described in more detail. The 5x CaCl2 solution is 40mM. You may be able to improve your results by using ice-cold 1x (8mM) CaCl2 and adding it drop-wise to the solution. The GRP78 protein is a ER marker but it will mark both smooth and rough ER, so the fact that there is plenty of GRP78 in your solution does not have to be an indication that the extraction was not complete. There are just smooth ER vesicles in the solution, which won't get precipitated until you spin it down at min 90 000xg.

A: With regard to your inquiry, my lab thinks that if the lysosomes are the same density and size as the ER population, they will be present in the ER collection. They recommend additional homogenization/lysis time on your cell population at the beginning of your enrichment process. They also think that further dilution could help as well. If possible, also try to add another centrifuge step to help remove other organelles prior to the high speed spin. We think these suggestions should help to eliminate the presence of the lysosomes in your ER collection.

A: We have evaluated this Kit in tissues. We have not yet tested this Kit in cell lines. However, this kit has been described for use with cell lines in citation #2 in the product manual.

A: We use the Wheaton grinder loose version. With our centrifuge we set the speed at 90000 g, our rotor doesn't have options to specify max or average settings, there is just a setting. But if the options on your rotor are for max or average, then I would imagine average would be the one to choose, but either one might work just as well.

A: We have used the kit with tissue. We haven't used the kit with cells. The customer will need to empirically determine the amount of cells needed. We unfortunately do not have any information on whether or not detergents will affect the usefulness of this kit. This kit is designed for endoplasmic reticulum enrichment. It is not designed for preservation of the Golgi and cell membrane fractions. The kit has been developed based on information in the general references listed in the manual provided with the kit.

A: The kit components for our kit NBP2-29482 do not contain any ascorbic acid. The only component that contains any acids is KC-121, and this component contains 1% acetic acid.

A: The short answer is Yes, this kit NBP2-29482 (Endoplasmic Reticulum Enrichment Extraction Kit) can be used in all cell lines. The yielding information/rate can also be obtained on our web datasheet. Endoplasmic Reticulum Enrichment Extraction Kit (NBP2-29482) datasheet, under the Title of "Applications/Dilutions".

A:

Our kit NBP2-29482 does not contain any polyclonal antibodies. The components list for this kit can be found on page five of the manual

A: We recommend you to review paper entitled "A rapid calcium precipitation method of recovering large amounts of highly pure hepatocyte rough endoplasmic reticulum" by Hamilton et al ( published in Journal of Lipid Research Volume 40, 1999) wherein the modified calcium precipitation method had been described in more detail. The 5x CaCl2 solution is 40mM. You may be able to improve your results by using ice-cold 1x (8mM) CaCl2 and adding it drop-wise to the solution. The GRP78 protein is a ER marker but it will mark both smooth and rough ER, so the fact that there is plenty of GRP78 in your solution does not have to be an indication that the extraction was not complete. There are just smooth ER vesicles in the solution, which won't get precipitated until you spin it down at min 90 000xg.

A: With regard to your inquiry, my lab thinks that if the lysosomes are the same density and size as the ER population, they will be present in the ER collection. They recommend additional homogenization/lysis time on your cell population at the beginning of your enrichment process. They also think that further dilution could help as well. If possible, also try to add another centrifuge step to help remove other organelles prior to the high speed spin. We think these suggestions should help to eliminate the presence of the lysosomes in your ER collection.

A: We have evaluated this Kit in tissues. We have not yet tested this Kit in cell lines. However, this kit has been described for use with cell lines in citation #2 in the product manual.

A: We use the Wheaton grinder loose version. With our centrifuge we set the speed at 90000 g, our rotor doesn't have options to specify max or average settings, there is just a setting. But if the options on your rotor are for max or average, then I would imagine average would be the one to choose, but either one might work just as well.

A: We have used the kit with tissue. We haven't used the kit with cells. The customer will need to empirically determine the amount of cells needed. We unfortunately do not have any information on whether or not detergents will affect the usefulness of this kit. This kit is designed for endoplasmic reticulum enrichment. It is not designed for preservation of the Golgi and cell membrane fractions. The kit has been developed based on information in the general references listed in the manual provided with the kit.

A: The kit components for our kit NBP2-29482 do not contain any ascorbic acid. The only component that contains any acids is KC-121, and this component contains 1% acetic acid.

A: The short answer is Yes, this kit NBP2-29482 (Endoplasmic Reticulum Enrichment Extraction Kit) can be used in all cell lines. The yielding information/rate can also be obtained on our web datasheet. Endoplasmic Reticulum Enrichment Extraction Kit (NBP2-29482) datasheet, under the Title of "Applications/Dilutions".

A:

Our kit NBP2-29482 does not contain any polyclonal antibodies. The components list for this kit can be found on page five of the manual

A: We recommend you to review paper entitled "A rapid calcium precipitation method of recovering large amounts of highly pure hepatocyte rough endoplasmic reticulum" by Hamilton et al ( published in Journal of Lipid Research Volume 40, 1999) wherein the modified calcium precipitation method had been described in more detail. The 5x CaCl2 solution is 40mM. You may be able to improve your results by using ice-cold 1x (8mM) CaCl2 and adding it drop-wise to the solution. The GRP78 protein is a ER marker but it will mark both smooth and rough ER, so the fact that there is plenty of GRP78 in your solution does not have to be an indication that the extraction was not complete. There are just smooth ER vesicles in the solution, which won't get precipitated until you spin it down at min 90 000xg.

A: With regard to your inquiry, my lab thinks that if the lysosomes are the same density and size as the ER population, they will be present in the ER collection. They recommend additional homogenization/lysis time on your cell population at the beginning of your enrichment process. They also think that further dilution could help as well. If possible, also try to add another centrifuge step to help remove other organelles prior to the high speed spin. We think these suggestions should help to eliminate the presence of the lysosomes in your ER collection.

A: We have evaluated this Kit in tissues. We have not yet tested this Kit in cell lines. However, this kit has been described for use with cell lines in citation #2 in the product manual.

A: We use the Wheaton grinder loose version. With our centrifuge we set the speed at 90000 g, our rotor doesn't have options to specify max or average settings, there is just a setting. But if the options on your rotor are for max or average, then I would imagine average would be the one to choose, but either one might work just as well.

A: We have used the kit with tissue. We haven't used the kit with cells. The customer will need to empirically determine the amount of cells needed. We unfortunately do not have any information on whether or not detergents will affect the usefulness of this kit. This kit is designed for endoplasmic reticulum enrichment. It is not designed for preservation of the Golgi and cell membrane fractions. The kit has been developed based on information in the general references listed in the manual provided with the kit.

A: The kit components for our kit NBP2-29482 do not contain any ascorbic acid. The only component that contains any acids is KC-121, and this component contains 1% acetic acid.

A: The short answer is Yes, this kit NBP2-29482 (Endoplasmic Reticulum Enrichment Extraction Kit) can be used in all cell lines. The yielding information/rate can also be obtained on our web datasheet. Endoplasmic Reticulum Enrichment Extraction Kit (NBP2-29482) datasheet, under the Title of "Applications/Dilutions".

A:

Our kit NBP2-29482 does not contain any polyclonal antibodies. The components list for this kit can be found on page five of the manual

A: We recommend you to review paper entitled "A rapid calcium precipitation method of recovering large amounts of highly pure hepatocyte rough endoplasmic reticulum" by Hamilton et al ( published in Journal of Lipid Research Volume 40, 1999) wherein the modified calcium precipitation method had been described in more detail. The 5x CaCl2 solution is 40mM. You may be able to improve your results by using ice-cold 1x (8mM) CaCl2 and adding it drop-wise to the solution. The GRP78 protein is a ER marker but it will mark both smooth and rough ER, so the fact that there is plenty of GRP78 in your solution does not have to be an indication that the extraction was not complete. There are just smooth ER vesicles in the solution, which won't get precipitated until you spin it down at min 90 000xg.

A: With regard to your inquiry, my lab thinks that if the lysosomes are the same density and size as the ER population, they will be present in the ER collection. They recommend additional homogenization/lysis time on your cell population at the beginning of your enrichment process. They also think that further dilution could help as well. If possible, also try to add another centrifuge step to help remove other organelles prior to the high speed spin. We think these suggestions should help to eliminate the presence of the lysosomes in your ER collection.

A: We have evaluated this Kit in tissues. We have not yet tested this Kit in cell lines. However, this kit has been described for use with cell lines in citation #2 in the product manual.

A: We use the Wheaton grinder loose version. With our centrifuge we set the speed at 90000 g, our rotor doesn't have options to specify max or average settings, there is just a setting. But if the options on your rotor are for max or average, then I would imagine average would be the one to choose, but either one might work just as well.

A: We have used the kit with tissue. We haven't used the kit with cells. The customer will need to empirically determine the amount of cells needed. We unfortunately do not have any information on whether or not detergents will affect the usefulness of this kit. This kit is designed for endoplasmic reticulum enrichment. It is not designed for preservation of the Golgi and cell membrane fractions. The kit has been developed based on information in the general references listed in the manual provided with the kit.

A: The kit components for our kit NBP2-29482 do not contain any ascorbic acid. The only component that contains any acids is KC-121, and this component contains 1% acetic acid.

A: The short answer is Yes, this kit NBP2-29482 (Endoplasmic Reticulum Enrichment Extraction Kit) can be used in all cell lines. The yielding information/rate can also be obtained on our web datasheet. Endoplasmic Reticulum Enrichment Extraction Kit (NBP2-29482) datasheet, under the Title of "Applications/Dilutions".

A:

Our kit NBP2-29482 does not contain any polyclonal antibodies. The components list for this kit can be found on page five of the manual

A: We recommend you to review paper entitled "A rapid calcium precipitation method of recovering large amounts of highly pure hepatocyte rough endoplasmic reticulum" by Hamilton et al ( published in Journal of Lipid Research Volume 40, 1999) wherein the modified calcium precipitation method had been described in more detail. The 5x CaCl2 solution is 40mM. You may be able to improve your results by using ice-cold 1x (8mM) CaCl2 and adding it drop-wise to the solution. The GRP78 protein is a ER marker but it will mark both smooth and rough ER, so the fact that there is plenty of GRP78 in your solution does not have to be an indication that the extraction was not complete. There are just smooth ER vesicles in the solution, which won't get precipitated until you spin it down at min 90 000xg.

A: With regard to your inquiry, my lab thinks that if the lysosomes are the same density and size as the ER population, they will be present in the ER collection. They recommend additional homogenization/lysis time on your cell population at the beginning of your enrichment process. They also think that further dilution could help as well. If possible, also try to add another centrifuge step to help remove other organelles prior to the high speed spin. We think these suggestions should help to eliminate the presence of the lysosomes in your ER collection.

A: We have evaluated this Kit in tissues. We have not yet tested this Kit in cell lines. However, this kit has been described for use with cell lines in citation #2 in the product manual.

A: We use the Wheaton grinder loose version. With our centrifuge we set the speed at 90000 g, our rotor doesn't have options to specify max or average settings, there is just a setting. But if the options on your rotor are for max or average, then I would imagine average would be the one to choose, but either one might work just as well.

A: We have used the kit with tissue. We haven't used the kit with cells. The customer will need to empirically determine the amount of cells needed. We unfortunately do not have any information on whether or not detergents will affect the usefulness of this kit. This kit is designed for endoplasmic reticulum enrichment. It is not designed for preservation of the Golgi and cell membrane fractions. The kit has been developed based on information in the general references listed in the manual provided with the kit.