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To the technical service,
we used with sucess in a previous work (Doublier et al.
BMC Cancer 2012) the antibody anti-HIF-1 alpha ChIP grade
(ab2185, lot GR-42498-2), bought in june 2011. We just
ordered and received another sample (lot GR-70784-1) of
this antibody and encountered the following problem: The
datasheet provided with the antibody bought last june
indicated in Application notes to use 25 microlliters of
ab2185 for 15 millions cells for ChIP assay. The datasheet
provided this time indicated in the same section 25
microliters of ab2185 for one million cells. We wonder if
this could be an error and if not, this sample of antibody
will be sufficient only for few experimental points. Could
you please clarify this point?
Thank you very much for your collaboration.
Asked on Apr 11 2012
Thank you for your enquiry regarding ab2185 - Anti-HIF1 alpha antibody - ChIP Grade.
I can confirm that ab2185 is a polyclonal antibody and not affinity purified; it is whole antiserum. As you may be aware there is usually batch-to-batch variations for polyclonals (vs monoclonals) depending on the immune response of the host animals (rabbit). I would advise you to use the recommended cell numbers indicated on the product datasheet which is regularly updated with the most up-to-date information.
I hope this will be useful for you. Should you still have any question with this antibody, then please do not hesitate to contact our Technical Department again.
Answered on Apr 11 2012