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Western after ChIP - how to do that?

ChiP-western

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#1 uhsiracu

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Posted 17 July 2013 - 04:05 AM

Hi all,
I need to do a western after the chip assay: I've already found some tips here in the forum, but I don't understand very well if I have to elute during the IP with the elution buffer or if I can elute with the loading buffer that I will use to load my sample in the SDS-PAGE gel. I'm using magnetic beads to bind the Fc of my ab. Can someone explain me better, please?
Thanks in advance!

#2 pcrman

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Posted 19 July 2013 - 03:59 PM

There is a previous discussion on this topic http://www.protocol-...osts/30800.html

Here is what we do for ChIP-Western: After chromatin immunoprecipitation with Protein G Dynabeads,, resuspend the beads in 2´ SDS sample buffer and boil for 5 min. Then collect the supernatant and proceed with western.

#3 uhsiracu

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Posted 20 July 2013 - 11:46 PM

Thanks for the reply. Another question: can I freeze the sample after boiling, or will freezing damage it?




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