Author: Timothy A. Dinh; Ramja Sritharan; F. Donelson Smith; Adam B. Francisco; Rosanna K. Ma; Rodica P. Bunaciu; Matt Kanke; Charles G. Danko; Andrew P. Massa; John D. Scott; Praveen Sethupathy
Title: Hotspots of aberrant enhancer activity in fibrolamellar carcinoma reveal molecular mechanisms of oncogenesis and intrinsic drug resistance Document date: 2020_1_18
ID: bf4qpsy7_78
Snippet: Protein lysates were prepared using RIPA buffer (Sigma, St. Louis, MO) supplemented with protease inhibitor cocktail (Sigma), phosphatase inhibitor cocktails 1 and 2 (Sigma), 1 mmol/L phenylmethylsulfonyl fluoride, 0.1% βmercaptoethanol, and 1 mmol/L dithiothreitol. Protein concentration was measured using the Pierce BCA Protein Assay (Thermo Fisher Scientific) according to the manufacturer's protocol. Lysates were subjected to sodium dodecyl su.....
Document: Protein lysates were prepared using RIPA buffer (Sigma, St. Louis, MO) supplemented with protease inhibitor cocktail (Sigma), phosphatase inhibitor cocktails 1 and 2 (Sigma), 1 mmol/L phenylmethylsulfonyl fluoride, 0.1% βmercaptoethanol, and 1 mmol/L dithiothreitol. Protein concentration was measured using the Pierce BCA Protein Assay (Thermo Fisher Scientific) according to the manufacturer's protocol. Lysates were subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) using 25 µg of lysate per lane under denaturing conditions in NuPAGE 10% Bis-Tris (Thermo Fisher Scientific) or homemade 12% Bis-Tris gels and transferred to PVDF membranes using a standard wet transfer protocol. Membranes were blocked with 5% dry nonfat milk in TBST and were probed with antibodies. Enhanced chemiluminescence reagent (GE Healthcare, Chicago, IL) was used for detection.
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