ChIP assay using Matrigel-3D cell culture - (Oct/23/2007 )

Hi, guys,

I will be doing ChIP from differentiated cell culture, which are growing in Matrigel.
By RIPA extraction, PBS/EDTA are used to dissolve Matrigel, but for ChIP, so questions come up are:

1. can formaldehyde-fixing also work for Matrigel-3D cell culture? i guess yes, since formaldehyde can probably go through gel and fix cells. is this correct?
2. cell harvesting is also like RIPA extraction?

thanks a lot and pls give me any idea of that.

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I'm sure the formaldehyde won't have any problem getting to your cells (it's a very small molecule). One issue I could see would be that your cells and the matrigel will all be crosslinked together and, after crosslinking, enzymes might not be very efficient at breaking the matrigel down. As long as the matrigel doesn't interfere with shearing this won't be a problem, but if it does, you might have to do some homogenizing (e.g. Tissue Tearor) prior to shearing.

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thanks for your post.

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