Anti-HSD3B1抗体[3C11-D4] (ab55268)

Anti-HSD3B1 antibody (ab55268) is raised against the full length protein, corresponding to amino acids 1-374 of Human HSD3B1.

Sequence alignment of Human HSD3B1 against HSD3B2 is 94%.

This suggests that ab55268 is not specific to HSD3B1 and will cross react with HSD3B2.  As the sequence homology is so similar, it may difficult to find an antibody specific to HSD3B1.

Thank you for your reply.

We recommend using 5% milk or BSA for the blocking step, however, when incubating with the primary antibody we recommend reducing that concentration to 1% or less. Blocking agent is not strictly necessary in the antibody diluent and can reduce the signal when used at higher concentrations.

I hope that this will help to improve our customer's results. If not, please let me know and I will be happy to offer either a free of charge replacement or a credit.

Thank you for contacting us and for forwarding the customer's protocol information. I am sorry that this antibody is not producing satisfactory results in WB. Could you please provide some additional information about the experiment?

1) Were the samples used human? Could you please let me know what samples are present in each lane of the first image using ab55268? It appears that there may be a band in the last lane.

2) When the customer reports using BSA and milk at 5%, was that 5% BSA and 5% milk in the same blocking buffer solution, or did the customer try each separately? Was the percentage also 5% for the antibody diluent? For the antibody diluent I typically recommend a blocking agent concentration less than 1%. High concentrations of other proteins can interfere with antibody binding and cause a loss of signal.

3) Could you please confirm the PO number or order reference number, or let me know what date the antibody was purchased? I have been unable to locate any orders of this antibody from your company.

I hope this will help to improve the customer's results. If not, please let me know and I will be happy to help you further.

Thank you for filling in the questionnaire and for reporting this problem to us. We will use the details you sent about your protocol in order to investigate the source of the problem. I have looked through the protocol, which looks fine to me. I appreciate the time that has been spent on these experiments and would be pleased to arrange a a credit note or a replacement with the alternative Mouse monoclonal HSD3B1 antibody ab69807 that has been tested in IHC-P and Human. The link to the datasheet of ab69807 is www.abcam.com/ab69807 I look forward to hearing from you with details of how you would like to proceed.

Thank you for contacting us. The HSD3B1 protein has 71% similarity between human and mouse. Theoretically the antibody should not be detecting the mouse protein however because we have not tested this so will not be able to confirm it. http://www.ebi.ac.uk/Tools/services/web/toolresult.ebi?jobId=clustalw2-I20110823-120040-0090-47470728-pg&tool=clustalw2&analysis=alignments If customer is happy using this ab with mouse lysates please let me know, I will be happy to send them an Abreview testing discount code. I would recommend using human lysates as positive control along with mouse lysates. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.