Western blot and immunoprecipitation
This protocol is extracted from research article:
STK38 promotes ATM activation by acting as a reader of histone H4 ufmylation
Sci Adv, Jun 3, 2020; DOI: 10.1126/sciadv.aax8214

Western blot and immunoprecipitation were performed as described previously (43). Cells were lysed with NETN buffer [20 mM tris-HCl (pH 8.0), 100 mM NaCl, 1 mM EDTA, and 0.5% NP-40] with 50 mM β-glycerophosphate, 10 mM NaF, and 1 mg/ml each of pepstatin A and aprotinin. The cell lysates were incubated with Benzonase to remove DNA and then incubated with antibody against protein of interest and protein A or protein G Sepharose beads (Amersham Biosciences) for 2 hours or overnight at 4°C. The immunoprecipitates were analyzed by SDS–polyacrylamide gel electrophoresis (SDS-PAGE) gels. Western blots were carried out following standard procedures.

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