Protease inhibitor cocktail 78410

Proteins were isolated from whole cells in lysis buffer (20mM Tris-HCl [pH 8], 137mM NaCl, 10% glycerol, 1% Triton X-100, and 2mM EDTA) containing Protease Inhibitor Cocktail 78410 (Thermo Fisher Scientific). Proteins were resolved by polyacrylamide gel electrophoresis on 10% sodium dodecyl sulfate gels EC60752 (Thermo Fisher Scientific) and immobilized onto nitrocellulose membranes (GE Healthcare, Chicago, IL, USA) by electroblot (Bio-Rad Laboratories, Hercules, CA, USA). Specific proteins were detected by immunoblotting with the indicated antibodies (STable 1); immunoreactive proteins were detected by enhanced chemiluminesence (Thermo Scientific) on a ChemiDoc XRS+ (Bio-Rad Laboratories).

Protein was extracted from the cells by treatment with lysis buffer
(20mM Tris-HCl [pH 8], 137mM NaCl, 10% glycerol, 1% Triton X-100, and 2mM EDTA)
containing Protease Inhibitor Cocktail 78410 (Thermo Scientific). Following
quantification by Bradford assay, the resulting lysate was resolved by
polyacrylamide gel electrophoresis with 10% sodium dodecyl sulfate gels EC60752
(Life Technologies) and immobilized onto nitrocellulose membranes (GE
Healthcare) by electroblot (Bio-Rad Laboratories). Membranes were probed for
KRT5 with MA5–12596, KRT14 with MA5–11599 and for β-actin
with MA5–15739 (Thermo Scientific). Blots were imaged using enhanced
chemiluminescence (Thermo Scientific) on a ChemiDoc XRS+ (Bio-Rad
Laboratories).