Anti-ATF6 抗体 (ab37149)

Thank you very much for your email.

We are very sorry to hear that you have problems with our antibodies. I will continue to investigate this issue with you. Please be assured, should we find that any of these antibodies would not work as expected, we will take the necessary actions. We do also guarantee all our products, and will replace the faulty product to you.

I have not found any indication that we would have sent a wrong antibody to you. The epitope of the monoclonal antibody ab11909is not known. That antibody was made against a full-length recombinant ATF6 protein and theepitope has not been mapped to our knowledge. We are also not aware of any specificity issues with this antibody, it has been tested against 3T3 cells in Western blot. We know that this antibody recognizes both full length and active/Cleaved ATF6.

The polyclonal antibody ab37149 has been raised against an immunogen derived from N terminus. It detects the alpha isoform.Due to the immunogen location it isalso predicted to to recognize the 50kD fragment.

I am not sure what the exact expected localisation of ATF6 should be? I have found this publication, however I am not sure how this relates to your results: "In 3-month-old lenses, ATF6 expression is still detectable in lens epithelium and fiber cells hthttp://www.sciencedirect.com/science/article/pii/S1567133X10001110"

Which antibody do you think provides you with the more specific staining? I can also offer to help to troubleshoot the protocol. Have you made the isotype controls also for each of the antibody - would there be anything technical we could improve? I would be pleased to look through the protocols as well. If you wish so, please do fill the attached questionnaire and send it back with an image of the results. This will then allow me to investigate this issue.

I look forward to hear back from you with your thoughts on this. Thank you for your cooperation.

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.



I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with one unit of ab65838,the order number is *****.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Thank you very much for your follow up email. I am sorry, that it took me so long to look into this.

Can you please comment on the following points:

1.) The expected Golgi and ER staining of the ab11909 could maybe not be observed in not very actively cycling cells. I would expect, that this signal would however be detected in ER and Golgi in cancer cells, such as the MCF7 cells. You might could do a positive control on such a cancer cell line.

2.) In regards to the ab37149, maybe there was a problem with this antibody vial. Although we do not have any indications that there would have been any problems, I can offer you to send you an alternative antibody for free. (and we would appreciate ifyoucould then provide us also with feedbacks on these via abreviews, https://www.abcam.com/abreviews) I could send you a free unit of ab65838, which is also against the N terminal and which therefore should provide a similar staining. The ab65838 has not been tested on frozen sections however. The ab65838 has been tested in Western blot and paraffin embedded tissue sections. It is very possible to work on frozen sections, however this needs to be tested experimentally first.

Please let me know what you think about this offer. I look forward hearing back from you.

Thank you for your reply. Unfortunately we do not know of a suitable positive control for the 50kDa band and have yet to test the antibody for the cleavage fragment. I hope that the recommendations I sent to you regarding protocol changes will help you detect the 50kDa band (use of a RIPA buffer, boil samples in loading buffer at 95-100C for 5 minutes, changing the blocking step, incubating the primary antibody overnight at 4C more concentrated, use Ecl+ detection kit and check the secondary antibody). Good luck with your experiments,