Western after ChIP - how to do that? - (Jul/17/2013 )
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!
There is a previous discussion on this topic http://www.protocol-online.org/biology-forums/posts/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.
Thanks for the reply. Another question: can I freeze the sample after boiling, or will freezing damage it?