I am going to do the co-immunoprecipitation of nuclear proteins.
To do extract nuclear protein in cell lysis, I need to use lysis buffer with higher ionic strength (e.g. 400mM of NaCl). Will it be a problem for the co-immunoprecipitation?
Will the protein interaction be disturbed by the buffer? And how to solve it?
Stéphane Roche
When you have your crude nuclear extract, the nuclei lysis buffer might be modify. For some experiment, I use Tris pH8 50mM, EDTA 10mM and SDS 1%. A second and classical buffer is 20 mM Tris HCl pH 8, 137 mM NaCl, 1% NP-40, 2 mM EDTA.
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