Ferroportin/SLC40A1 Antibody - BSA Free

A: Optimization could really make a difference as we have seen some staining with the same antibody on frozen section. The main thing would be to focus on antigen retrieval step and the buffer used. For the antigen retrieval method, we would suggest an increase to at least 10-20 minutes to the pressure cooker step of the HIER method; the duration of the retrieval step can greatly determine if the antigen will be exposed or not for the antigen binding site of the antibody to recognize the protein. Also, I the incubation time for peroxidase blocking time could be reduced from 30min to 15 min, as we have mentioned. Since this is a TMP, permeabilization reagents should be used for the experiment. You can review what we have available on our site in terms of our general protocols:HIER and IHC-P

A: NBP1-21502 is still the same antibody it was two years ago. Mouse was mistakenly removed during a website update and has since been re-added. This antibody still cross-reacts with mouse. In the past two years we have begun to validate more of our products for use in ICC and this is one that has had its uses updated since you last purchased.

A: The immunogen used for this Ferroportin/SLC40A1 Antibody NBP1-21502 falls within a helical domain of the protein and some permeabilization is recommended for staining. Our lab uses 0.2% Tween-20 included in the blocking and diluent buffers to perform this permeabilization. As our directly conjugated antibodies are prepared per order, we unfortunately do not have sample sizes available.

A: There is a western blot protocol specific to this antibody that can be found on the datasheet; it differs from your protocol quite a lot. I would give this protocol a try and see if you get better results.Although I am uncertain of your blocking method, we find that BSA and PVDF can give high non-specific background so the nitrocellulose and NFDM are recommend for this antibody. In addition, there seems to be no protein expression of the SLC40A1 in liver or spleen based on Human Protein Atlas.https://www.proteinatlas.org/ENSG00000138449-SLC40A1/tissueA good positive control might be bone marrow.

A: With regards to conjugating NBP1-21502SS (0.025 ml) to PE or PerCP, there are two possibilities: The first is that you could conjugate the antibody yourself, using easy-to-use kits. You would choose from Lightning-Link R-PE 3 x 0.01 mg, or Lightning-Link PerCP 3 x 0.01 mg. Because the buffer of the NBP1-21502 antibody is Tris-glycine, which is not compatible with the Lightning-Link conjugation chemistry as that targets amine groups, you would perform a buffer exchange first using the Antibody Concentration and Clean Up Kit. The lab has informed me that yes, they could conjugate your choice of PE or PerCP to this antibody for you. However, it would have to be the full-size antibody vial (0.1 ml) rather than the sample size (0.025 ml).

A: Optimization could really make a difference as we have seen some staining with the same antibody on frozen section. The main thing would be to focus on antigen retrieval step and the buffer used. For the antigen retrieval method, we would suggest an increase to at least 10-20 minutes to the pressure cooker step of the HIER method; the duration of the retrieval step can greatly determine if the antigen will be exposed or not for the antigen binding site of the antibody to recognize the protein. Also, I the incubation time for peroxidase blocking time could be reduced from 30min to 15 min, as we have mentioned. Since this is a TMP, permeabilization reagents should be used for the experiment. You can review what we have available on our site in terms of our general protocols:HIER and IHC-P

A: NBP1-21502 is still the same antibody it was two years ago. Mouse was mistakenly removed during a website update and has since been re-added. This antibody still cross-reacts with mouse. In the past two years we have begun to validate more of our products for use in ICC and this is one that has had its uses updated since you last purchased.

A: The immunogen used for this Ferroportin/SLC40A1 Antibody NBP1-21502 falls within a helical domain of the protein and some permeabilization is recommended for staining. Our lab uses 0.2% Tween-20 included in the blocking and diluent buffers to perform this permeabilization. As our directly conjugated antibodies are prepared per order, we unfortunately do not have sample sizes available.

A: There is a western blot protocol specific to this antibody that can be found on the datasheet; it differs from your protocol quite a lot. I would give this protocol a try and see if you get better results.Although I am uncertain of your blocking method, we find that BSA and PVDF can give high non-specific background so the nitrocellulose and NFDM are recommend for this antibody. In addition, there seems to be no protein expression of the SLC40A1 in liver or spleen based on Human Protein Atlas.https://www.proteinatlas.org/ENSG00000138449-SLC40A1/tissueA good positive control might be bone marrow.

A: With regards to conjugating NBP1-21502SS (0.025 ml) to PE or PerCP, there are two possibilities: The first is that you could conjugate the antibody yourself, using easy-to-use kits. You would choose from Lightning-Link R-PE 3 x 0.01 mg, or Lightning-Link PerCP 3 x 0.01 mg. Because the buffer of the NBP1-21502 antibody is Tris-glycine, which is not compatible with the Lightning-Link conjugation chemistry as that targets amine groups, you would perform a buffer exchange first using the Antibody Concentration and Clean Up Kit. The lab has informed me that yes, they could conjugate your choice of PE or PerCP to this antibody for you. However, it would have to be the full-size antibody vial (0.1 ml) rather than the sample size (0.025 ml).

A: Optimization could really make a difference as we have seen some staining with the same antibody on frozen section. The main thing would be to focus on antigen retrieval step and the buffer used. For the antigen retrieval method, we would suggest an increase to at least 10-20 minutes to the pressure cooker step of the HIER method; the duration of the retrieval step can greatly determine if the antigen will be exposed or not for the antigen binding site of the antibody to recognize the protein. Also, I the incubation time for peroxidase blocking time could be reduced from 30min to 15 min, as we have mentioned. Since this is a TMP, permeabilization reagents should be used for the experiment. You can review what we have available on our site in terms of our general protocols:HIER and IHC-P

A: NBP1-21502 is still the same antibody it was two years ago. Mouse was mistakenly removed during a website update and has since been re-added. This antibody still cross-reacts with mouse. In the past two years we have begun to validate more of our products for use in ICC and this is one that has had its uses updated since you last purchased.

A: The immunogen used for this Ferroportin/SLC40A1 Antibody NBP1-21502 falls within a helical domain of the protein and some permeabilization is recommended for staining. Our lab uses 0.2% Tween-20 included in the blocking and diluent buffers to perform this permeabilization. As our directly conjugated antibodies are prepared per order, we unfortunately do not have sample sizes available.

A: There is a western blot protocol specific to this antibody that can be found on the datasheet; it differs from your protocol quite a lot. I would give this protocol a try and see if you get better results.Although I am uncertain of your blocking method, we find that BSA and PVDF can give high non-specific background so the nitrocellulose and NFDM are recommend for this antibody. In addition, there seems to be no protein expression of the SLC40A1 in liver or spleen based on Human Protein Atlas.https://www.proteinatlas.org/ENSG00000138449-SLC40A1/tissueA good positive control might be bone marrow.

A: With regards to conjugating NBP1-21502SS (0.025 ml) to PE or PerCP, there are two possibilities: The first is that you could conjugate the antibody yourself, using easy-to-use kits. You would choose from Lightning-Link R-PE 3 x 0.01 mg, or Lightning-Link PerCP 3 x 0.01 mg. Because the buffer of the NBP1-21502 antibody is Tris-glycine, which is not compatible with the Lightning-Link conjugation chemistry as that targets amine groups, you would perform a buffer exchange first using the Antibody Concentration and Clean Up Kit. The lab has informed me that yes, they could conjugate your choice of PE or PerCP to this antibody for you. However, it would have to be the full-size antibody vial (0.1 ml) rather than the sample size (0.025 ml).

A: Optimization could really make a difference as we have seen some staining with the same antibody on frozen section. The main thing would be to focus on antigen retrieval step and the buffer used. For the antigen retrieval method, we would suggest an increase to at least 10-20 minutes to the pressure cooker step of the HIER method; the duration of the retrieval step can greatly determine if the antigen will be exposed or not for the antigen binding site of the antibody to recognize the protein. Also, I the incubation time for peroxidase blocking time could be reduced from 30min to 15 min, as we have mentioned. Since this is a TMP, permeabilization reagents should be used for the experiment. You can review what we have available on our site in terms of our general protocols:HIER and IHC-P

A: NBP1-21502 is still the same antibody it was two years ago. Mouse was mistakenly removed during a website update and has since been re-added. This antibody still cross-reacts with mouse. In the past two years we have begun to validate more of our products for use in ICC and this is one that has had its uses updated since you last purchased.

A: The immunogen used for this Ferroportin/SLC40A1 Antibody NBP1-21502 falls within a helical domain of the protein and some permeabilization is recommended for staining. Our lab uses 0.2% Tween-20 included in the blocking and diluent buffers to perform this permeabilization. As our directly conjugated antibodies are prepared per order, we unfortunately do not have sample sizes available.

A: There is a western blot protocol specific to this antibody that can be found on the datasheet; it differs from your protocol quite a lot. I would give this protocol a try and see if you get better results.Although I am uncertain of your blocking method, we find that BSA and PVDF can give high non-specific background so the nitrocellulose and NFDM are recommend for this antibody. In addition, there seems to be no protein expression of the SLC40A1 in liver or spleen based on Human Protein Atlas.https://www.proteinatlas.org/ENSG00000138449-SLC40A1/tissueA good positive control might be bone marrow.

A: With regards to conjugating NBP1-21502SS (0.025 ml) to PE or PerCP, there are two possibilities: The first is that you could conjugate the antibody yourself, using easy-to-use kits. You would choose from Lightning-Link R-PE 3 x 0.01 mg, or Lightning-Link PerCP 3 x 0.01 mg. Because the buffer of the NBP1-21502 antibody is Tris-glycine, which is not compatible with the Lightning-Link conjugation chemistry as that targets amine groups, you would perform a buffer exchange first using the Antibody Concentration and Clean Up Kit. The lab has informed me that yes, they could conjugate your choice of PE or PerCP to this antibody for you. However, it would have to be the full-size antibody vial (0.1 ml) rather than the sample size (0.025 ml).

A: Optimization could really make a difference as we have seen some staining with the same antibody on frozen section. The main thing would be to focus on antigen retrieval step and the buffer used. For the antigen retrieval method, we would suggest an increase to at least 10-20 minutes to the pressure cooker step of the HIER method; the duration of the retrieval step can greatly determine if the antigen will be exposed or not for the antigen binding site of the antibody to recognize the protein. Also, I the incubation time for peroxidase blocking time could be reduced from 30min to 15 min, as we have mentioned. Since this is a TMP, permeabilization reagents should be used for the experiment. You can review what we have available on our site in terms of our general protocols:HIER and IHC-P

A: NBP1-21502 is still the same antibody it was two years ago. Mouse was mistakenly removed during a website update and has since been re-added. This antibody still cross-reacts with mouse. In the past two years we have begun to validate more of our products for use in ICC and this is one that has had its uses updated since you last purchased.

A: The immunogen used for this Ferroportin/SLC40A1 Antibody NBP1-21502 falls within a helical domain of the protein and some permeabilization is recommended for staining. Our lab uses 0.2% Tween-20 included in the blocking and diluent buffers to perform this permeabilization. As our directly conjugated antibodies are prepared per order, we unfortunately do not have sample sizes available.

A: There is a western blot protocol specific to this antibody that can be found on the datasheet; it differs from your protocol quite a lot. I would give this protocol a try and see if you get better results.Although I am uncertain of your blocking method, we find that BSA and PVDF can give high non-specific background so the nitrocellulose and NFDM are recommend for this antibody. In addition, there seems to be no protein expression of the SLC40A1 in liver or spleen based on Human Protein Atlas.https://www.proteinatlas.org/ENSG00000138449-SLC40A1/tissueA good positive control might be bone marrow.

A: With regards to conjugating NBP1-21502SS (0.025 ml) to PE or PerCP, there are two possibilities: The first is that you could conjugate the antibody yourself, using easy-to-use kits. You would choose from Lightning-Link R-PE 3 x 0.01 mg, or Lightning-Link PerCP 3 x 0.01 mg. Because the buffer of the NBP1-21502 antibody is Tris-glycine, which is not compatible with the Lightning-Link conjugation chemistry as that targets amine groups, you would perform a buffer exchange first using the Antibody Concentration and Clean Up Kit. The lab has informed me that yes, they could conjugate your choice of PE or PerCP to this antibody for you. However, it would have to be the full-size antibody vial (0.1 ml) rather than the sample size (0.025 ml).