HCT116 cells seeded in a six-well plate were PBS treated or infected with viruses for 48 h, and the cells were harvested and lysed with RIPA lysis buffer containing protease and phosphatase inhibitors. Protein concentration was measured using the BCA protein assay kit (Thermo Fisher Scientific, Carlsbad, CA, USA), and equal amounts of proteins from each sample were separated using SDS-PAGE and transferred to polyvinylide difluoride membrane (Thermo Fisher Scientific). The antibodies against PARP, PKR, p38 MAPK, p-p38 MAPK (Thr180/Tyr182), JNK, p-JNKs (Thr183/Thr183/Thr221), STAT3, and p-STAT3 (Tyr705) were purchased from Cell Signaling Technology (Danvers, MA, USA), ERK1/2, p-ERK1 (Thr202/Tyr204)/ERK2 (Thr185/Tyr187), Bad, Bcl-xL, and Bcl-2 were purchased from Beyotime Institute of Biotechnology (China). β-Actin was obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Immunoreactive bands were visualized with chemiluminescence using ECL Western blot detection reagents (Santa Cruz Biotechnology).
Polyvinylide difluoride membrane
Manufactured by Thermo Fisher Scientific
Sourced in United States
Polyvinylide difluoride membrane is a type of lab equipment used for various applications in scientific research and analysis. It is a durable and chemically resistant material that can be used for techniques such as Western blotting, membrane-based assays, and sample preparation. The core function of this membrane is to act as a physical support and transfer medium for biomolecules during laboratory procedures.
Automatically generated - may contain errors
Lab products found in correlation
β actin, by Santa Cruz Biotechnology (2 mentions)
Ecl western blot detection reagents, by Santa Cruz Biotechnology (2 mentions)
P stat3 tyr705, by Cell Signaling Technology (1 mentions)
Bcl 2, by Beyotime (1 mentions)
Erk1 2, by Beyotime (1 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (1 mentions)
Bcl xl, by Beyotime (1 mentions)
Halt protease inhibitor cocktail, by Thermo Fisher Scientific (1 mentions)
β catenin, by Beyotime (1 mentions)
2 protocols using polyvinylide difluoride membrane
1
Evaluating Cellular Signaling Pathways
2
Virus-Induced Signaling Pathway Analysis
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Cells infected with virus or mock-infected cells were harvested and lysed with RIPA lysis buffer containing protease and phosphatase inhibitors (Halt Protease Inhibitor Cocktail, Thermo Fisher Scientific Inc.). Equal amounts of protein from each sample were subjected to electrophoresis on SDS-polyacrylamide gel and transferred to polyvinylide difluoride membrane (Thermo Scientific™, USA). Rabbit monoclonal antibody for PI3K (Cell Signaling Technology, Danvers, MA, USA), β-catenin (Beyotime Institute of Biotechnology, China), phospho-Akt (Ser473), Akt (Cell Signaling Technology, Danvers, MA, USA), phospho-GSK-3β (Ser9), GSK-3β (Beyotime Institute of Biotechnology, China), Bxl-xL (Beyotime Institute of Biotechnology, China) and mouse monoclonal antibodies for β-actin (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) were used as primary antibodies. Immunoreactive bands were visualized with chemiluminescence using ECL western blot detection reagents (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA).
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