Anti-acetyl Lysine antibody (ab76)

I am interested in performing Western blots to test for acetylated lysine residues in yeast proteins, including (but not exclusive to) the histone proteins. I looked at the acetyl-lysine antibodies your company produces, and I'm wondering which antibody you would recommend for this application. Has it been tested on yeast, and if not, would it be possible to get a trial size to test it out?

ANSWER:

Thank you for your enquiry.

This antibody is currently under evaluation for its use as a pan-acetyl-lysine antibody. However, the immunogen used to raise this antibody was a tetra-acetylated synthetic N-terminal peptide, corresponding to amino acids 1-18 of Human histone H4. It has been shown to react against histones, PCAF and E2F. Given the latter two it is likely to recognise acetylated non-histone proteins.

It has not been tested against yeast. However, given the conservation of the histone tails and the broad spectrum of specificity I consider this likely.

Should you require further assistance, please do not hesitate to contact me.

I dont know if this should be posted via your webpage or not? Nevertheless, here is our answers to your questions:

We have not tested ab6748 without any blocking agent and you are right this would also be a nice control to do. On the other hand we have obtained negative results with 4 different blocking agents and therefore we seriously believe that there is something wrong with this antibody. To our experience in is normal to have problems with maybe 1 or 2 blocking agents but not with 4 different.

No permeabilising agent was used.

As mentioned previously the delution buffer was TBS (Trizma Base-sodiumchloride pH 7,4. Trizma base 6,06g and Sodium chloride 8,76g / l)

We would appreciate to get this antibody (ab6748) to work together with a primary antibody also bought from Abcam (Anti-acetyl lysine ab76). We would therefore suggest that you supply us with the protocol originally used to test the ab6748 for IHC together with another batch of the ab6748 free of charge. We would then perform an experiment comparing the two batches of ab6748 and supply you with the result. This should be in your favour as well as there could be other customers having problems with the 67077 batch.

ANSWER:

Thank you for your enquiry.

My colleague Dr Sarah Mardle is out of office until Thursday morning. As soon as she gets back, she will get in touch with you and deal with your enquiry.

Thank you for your patience.

Have you tested the pan-specificity for this antibody. I want to use it for yeast extracts - is this antibody specific yeast histones as well.

ANSWER:

We have not further tested the cross-reactivity of this antibody, all we can say is that it is likely to cross react with yeast based on sequence homology.

I would expect it to work in yeast as there is a high level of histone homology between species.

I want to know whether you have monoclonal antibodies for the different isoforms of acetylated histone h4 and if you do, pls. send me the data sheetes for all of them including their prices and the quantities available.

Thanks very much, Samadhi Vitharana, University of Kansas.

ANSWER:

We do not currently have monoclonals to the different isoforms, but expect to add these to the range soon. Abcam