Anti-Amyloid beta precursor protein antibody [Y188] (ab32136)

I will try that out and let you know the results.Best,

ANSWER:

According to our records, ab32136 was proving difficult to use in IHC and we were in contact in order to help resolve the issue. Looking at our correspondence, it appears that we are awaiting more details in order to help us better understand the difficulties experienced. If the requested information has already been sent, it appears that it did not reach our Scientific Support team and we apologize for this inconvenience. In this case we would like to ask for the information again so that we can reach a resolution. If the issue has already been settled, please let us know so that we can be assured that the problem has been solved to your satisfaction and update our records. We wish you the best of luck with your research and look forward to a reply.

Please find the file attached for your information. Hope to hear from you soon. I expect to have a replacement. Thanks.

ANSWER:

Thank you for filling the questionnaire and for sending the images.

I have read the whole protocol which looks fine to me. However I have further suggestions to make which I am sure will help to improve results;

Antigen retrieval step; this is a crucial step in IHC-P; we always recommend our customer optimizing this step. If pressure cooker was used we recommend incubating sections for 3 minutes; Three minutes is only suggested as a starting point antigen retrieval time. Less than 3 minutes may leave the antigens under-retrieved, leading to weak staining. More than 3 minutes may leave them over-retrieved, leading to non-specific background staining and also increasing the chances of sections dissociating from the slides. A control experiment is recommended beforehand, where slides of the same tissue section are retrieved for 1, 2, 3, 4 and 5 minutes before being immunohistochemically stained to evaluate optimum antigen retrieval time for the particular antibody being used.

Although blocking buffer does not affect the specificity of antibody however under ideal conditions the blocking buffer species should be same as secondary antibody and we recommend blocking tissue sections with serum or BSA for 1-2 hours at RT.

I am sure with optimization in protocol the results will improve. If however these suggestions does not help please contact me back I will be happy to assist you further.