Hi,

A new collagen gel culture question.
Is it possible to perform western blotting after culturing cells in a collagen gel culture?
Something that you shall think about.

I have found some articles were they only have digested the collagen with collagenase before lysation of the cells, and then centrifugation before they run a western blot. But since I'm new I'm not totally sure if there are something "different" form regular protocols where the cells are cultured on plastic etc.

//Camilla

You might want to try out in-cell fluorescent Westerns - they work on two color channels (one with a reference protein), so they give you relative quantitation.
I've only done IF, and no Westerns with these, but do not see any problem following collagen digestion other than protein overloading.