Hi,
I am preparing cell lysates using NIH3T3 cells. However I am achieving very less protein concentration. The lysis buffer works well with other cells.
Can you share your experience with these cells?
Thank you,
Sumersing Patil
Hi Sebastian,
I don't know the exact number of cells I use at the time of lysate preparation. However, I use 80% confluent cells grown in T150 cell culture flask. For Lysis, I incubate the detached cells in lysis buffer (0.5 mL per flask ) for 40 mins on ice with intermittent vortexing. In this manner I was able to achieve 1mg/mL protein. Meaning, 0.5 mg protein per flask.
Thanks,
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