Anti-acetyl Lysine antibody (HRP) (ab23364)

I am really looking for the 'post-sales' department with respect to an antibody I recently purchased from you.

The acetyl-lysine antibody conjugated to HRP (ab23364) is certainly not specific to acetylated proteins when compared to an unconjugated acetyl lysine antibody from a reval company. I am unable to see the band that corresponds to acetylated histones in my samples. The antibody you supply shows no difference in banding patterns when compared between a hyper acetylated (treated) sample and an untreated sample.

Could you possibly help, as I am desperate for an acetyl-Lysine AB+HRP that works effectively?

ANSWER:

Thank you for your enquiry.

I am sorry to hear that you have been having problems with ab23364 anti-acetyl Lysine. Quality is important to Abcam, if an antibody does not work as specified on the datasheet, we will offer a replacement or reimbursement if the antibody was delivered within the past 90 days.

I have examined your complaint and I have a few comments. It is very difficult for me to comment on the performance of our antibody when compared to a competitor as I am unaware of the degree of cross reactivity the antibody may demonstrate against non-acetylated proteins or the number of acetylated proteins that it may recognise by western blot. Even if the immunising peptide was the same this is likely to vary from antiserum to antiserum.

You mentioned acetylated histones so I am assuming that these are your targets of interest. Please can I ask whether you have tried acid extracting histones with and without 5mM Sodium Butyrate treatment before performing western blotting. This will enable you to determine whether the increase in acetylation is detected by this antibody. To control this experiment it is essential the loadings are equalised by a parallel Coomassie stained histone prep or examination of the Ponceau transfer.

Abcam has an excellent histone extraction protocol located at the link below.

http://marketing.abcam.com/index.html?pageconfig=view_protocol&pid=316

Should you feel that the antibody is still not performing after considering my comments please submit a technical questionnaire by clicking on the link below.

http://www.abcam.com/index.html?section=western&pageconfig=technical&intAbID=23364&mode=questionaire

Please do not hesitate to contact me should you require further assistance.