Nuclear extraction buffer 2 (NEB2)
10 mM HEPES (pH 8)
Phosphatase inhibitor cocktail (PhosSTOP, Roche Applied Science) (optional; see note below)
Protease inhibitor cocktail (one tablet per 20 mL) (Complete, Mini; Roche Applied Science)
The protocol may require a second extraction with a lower concentration of Triton X-100.
If the protein of interest is not a kinase or kinase substrate, the phosphatase inhibitor cocktail can be omitted.
To adapt the above buffer for use in chromatin immunoprecipitation procedures, add the following reagents:
0.1 mg/mL 4-(2-aminoethyl)benzenesulfonyl fluoride hydrochloride (AEBSF)
