Hi,
I am going to perform brain clearing CUBIC protocol and before that I need to do western blot with the perfused brain tissue. Does anyone suggest me the protocol for the lysis of brain tissue, which buffer has be used. I have done western but not the initial lysis step and that too for Brain tissue. I am very new to this and would appreciate any suggestions.
Thanks in advance.
DearAshwini
I am agree with Sebastian
If your tissue fixed with formalin,you can not use them.
If not, these tissue can be easily use for homogenization and protein extraction. You must lysed the cells using urea and thiourea and concurrently use of protease inhibitor or work on ice under liquid nitrogen.
You can use any cell lysis buffer with protease and phosphatase inhibitors ( Lysis buffer D,CS, RIPA etc.,). It also depends on which protein you are looking for membrane/cytoskeletal/cytoplasmic ?
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