I am interested in performing a western blot on cells that I have grown in Matrigel. I have read different protocols on recovering the cells from the matrix, including use of ice-cold PBS, Corning Dispase, or some lysis buffer. However, my protein of interest is one that is secreted into the ECM, so I am interested in including any protein in the matrigel as well, and it seems most protocols involve separating the cells from the matrigel. How can I prep lysates that include my cells and the surrounding matrigel?
Add equal volumes of a boiling lysis buffer: 2% SDS, 2.0 mM sodium ortho-vanadate, 20 mM Tris pH 7.4). Transfer to a new tube using large orifice tubes. Boil in heat block x 5 minutes, then vortex, then boil again.
Alternatively, use less matrigel on the next experiment, which will allow it to liquefy easier.
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