Anti-KMT1E / SETDB1 antibody - ChIP Grade (ab12317)

How about the western image you are showing on the website?

Lane 1 and 2 were mock transfercted and 3 and 4 were SETDB1 expressed.

However, why are the bands of Lane 1 and 2 heavier than those of Lane 3 and

4?

ANSWER:

Thanks for your enquiry. There are two western blots on the datasheet for ab12317. I believe you are referring to the bottom image. I think there is a mistake in the annotation for this blot. 'Mock transfected' should refer to lanes 3 and 4 of this blot, not lanes 1 and 2. I am sorry for the confusion here. I hope this information helps to explain things. Please feel free to contact me again with any further questions.

Hi,

I purchased your antibody, ab12317, and did western blot. But there are two bands showing in my western.

Could you explain the image showing on your webpage? And which band should be the SETDB1?

If the band should be 155Kd, why what you are showing is more than 160Kd?

ANSWER:

Thank you for your enquiry. The predicted molecular weight of SETDB1 is 155kDa, based on the amino acid composition of the protein. It should be noted however that this protein is known to be phophorylated, as indicated on the Swissprot data page entry for SETDB1 under the 'Sequence annotation (Features)' section: http://www.uniprot.org/uniprot/Q15047 Phosphorylation of a protein will slow its migration through a gel, making it appear much heavier than normal. This may be what you are seeing. Alternatively, sometimes proteins run a little slower than expected based on their amino acid composition, even without post translational modifications. This may be what is happening here. Similar results are seen with ab12317, another antibody to SETDB1. ab12317 detects a band of 170kDa via western blotting. I hope this is helpful. Feel free to send me an image of your blot and I'll be happy to look at it.  Please contact me again if you have any further questions.

Customer's question: I would like to the sequence of SET DB1 protein to which antibody is raised. I am interested to buy a SETDB1 antibody which recognizes C terminus of my SETDB1 protein.

ANSWER:

The precise sequence used to generate ab12317 is proprietary.

The epitope recognized by ab12317 is found between residues 1125 and 1175 of SwissProt entry Q15047 (http://us.expasy.org/cgi-bin/niceprot.pl?Q15047).