Nuclear extraction buffer 1 (NEB1)

2 mM EDTA

2.5 mM DTT

10 mM HEPES (pH 8)

0.4 M sucrose

1 mM sodium vanadate (Na₃VO₄) (optional; see note below)

5 mM sodium fluoride (NaF) (optional; see note below)

Protease inhibitor cocktail (one tablet per 50 mL) (Complete, Mini; Roche Applied Science)

If the protein of interest is not a kinase or kinase substrate, NaF and Na₃VO₄ can be omitted.

To adapt the above buffer for use in chromatin immunoprecipitation procedures, omit the DTT and add the following reagents:

0.1 mg/mL 4-(2-aminoethyl)benzenesulfonyl fluoride hydrochloride (AEBSF)

5 mM β-mercaptoethanol