Hello,
I am planning on doing a TMT-labeled MS analysis experiment for global proteomics of glioma primary stem cells. Its my first time doing the experiment so Im referencing the protocols of my predecessors. But all of them used different lysis buffers.. one of the protocols use a 8M urea, 100mM TEAB buffer + In-solution digestion of liver tissue
another uses RIPA buffer + S-trap digestion method on a lung cancer cell line.
What does the buffer depend on? Does it depend on the type of digestion method? The type of tissue? the TMT itself?
any help would be appreciated,
Andrew