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Anti-SP1 (phospho T453) antibody - ChIP Grade (ab59257)

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1 question for ab59257

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Question 1

Wednesday 23-June-2010

I am looking into buying one of your Sp1 phosphoT453 antibody (ab37707 or ab59257). None of them have been reviewed for western blot which is what we want to use it for. I am confused by the size of products that you show on the western blot for ab37707.....In Hela cells (and most other cell types), Sp1 runs at 80-90Kda, (as shown with your other Sp1 antibody ab77441) yet the products recognized with that antibody appear much higher.

ANSWER:

  

SP1 has a number of glycosylation sites and so this is probably the reason why we see a difference between the expected MW (80-90kDa) (ab59257) and the observed (120kDa) in our product ab37707. We have run ab37707 alongside ab59257. No information was shown to suggest the 120kDa band was non-specific, but for ab59257 we observed a band in Jurkat cells comparable to the 120kDa band we see in ab37707.

There is also a difference between the target band in HeLa and Jurkat. We cannot predict which glycosylated form(s) will be present in the lysate used, and so this is likely to be the reason for the difference observed.

We have also carried out a peptide block, the data of which is on the datasheet and shows that the 120kDa is quenched by the immunising modified peptide and not by the non-modified peptide.

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