I need to obtain cell lysates in order to perform Western Blots to check the presence od MMP-2 and MMP-9. What would be the best lysis buffer?
I have used NP-40 and RIPA, they both work for western. If your cells are in culture then you can also check MMMP-2 and 9 from the conditioned media. Incubate cells for 12-24 hr in media without serum, collect media, mix with loading buffer, boil and run the gel. You can also use the c-media to run zymograms and check for activity of MMP-2 and 9.
We have used RIPA (medium strength) and we had excellent results for Western MMP-2 an 9. The c-media for zymography as suggested by Ariztia is worth trying as well
C-medium means conditioned medium. Thus, when you culture your cells in a medium with no or reduced (say 1% ) FBS (or the serum type you use). The resulting culture supernatant is a conditioned medium
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