Hello
I'm working on melanoma tumors from mouse and I want to extract proteins from it. I have a protocol where the detergent used is 10% Brij.
Is the concentration too high ? Or is it normal because it's from the tumor and not from cells ?
Thanks for your help
Hi Camille,
I always used the RIPA buffer with 1% Triton X-100 in the 1X lysis buffer. Both Brij and Triton X-100 are non-ionic detergent, thus I suggest you to add your compound at a lower concentration. 1% is suggested as final concentration in most of the lysis buffer recipes. To increase the ionic strength, add 1% sodium deoxycholate (final) in your lysis buffer. I don't think is a matter of the tissue you're working with. Usually, mouse melanoma tumors (e.g. from B16 cells) smashing easily just pipetting. Good luck!
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