Nuclear extraction buffer 3 (NEB3)

10 mM HEPES (pH 8)

2 mM MgCl₂

Phosphatase inhibitor cocktail (PhosSTOP, Roche Applied Science) (optional; see note below)

Protease inhibitor cocktail (one tablet per 10 mL) (Complete, Mini; Roche Applied Science)

1.7 M sucrose

0.15% (v/v) 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)

5 mM β-mercaptoethanol