Cells were washed with PBS and lysed in RIPA lysis buffer (Thermo Fisher Scientific) with Halt Protease Inhibitor Cocktail (Thermo Fisher Scientific) and Benzonase (Sigma-Aldrich) for 20 min on ice. The insoluble fraction was removed by centrifugation, the protein concentration was quantified using a BCA protein assay kit (Thermo Fisher Scientific), and an equal amount of lysate was run on SDS–PAGE 4–12% Bis–Tris Protein Gels (Thermo Fisher Scientific) and then transferred to nitrocellulose membrane with a XCell II Blot Module Wet Tank Transfer System (Thermo Fisher Scientific). Membranes were blocked in Intercept (PBS) Blocking Buffer (LI-COR Biosciences) and incubated with primary antibodies overnight at 4 °C. The membranes were then washed in Tris-buffered saline with Tween-20 (TBS-T), incubated for 1 h with secondary IRDye-conjugated antibodies (LI-COR Biosciences) and washed three times in TBS-T for 5 min before near-infrared western blot detection on an Odyssey Imaging System (LI-COR Biosciences).
Sds page 4 12 bis tris protein gels
Manufactured by Thermo Fisher Scientific
SDS–PAGE 4–12% Bis–Tris Protein Gels are laboratory equipment used for the separation and analysis of proteins based on their molecular weight. These gels provide a consistent and reliable platform for protein electrophoresis, allowing for the effective separation and visualization of complex protein samples.
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Lab products found in correlation
Bca protein assay kit, by Thermo Fisher Scientific (4 mentions)
Ripa lysis buffer, by Thermo Fisher Scientific (4 mentions)
Intercept pbs blocking buffer, by LI COR (2 mentions)
Odyssey imaging system, by LI COR (2 mentions)
Irdye conjugated antibody, by LI COR (2 mentions)
Xcell 2 blot module wet tank transfer system, by Thermo Fisher Scientific (2 mentions)
Halt protease inhibitor cocktail, by Thermo Fisher Scientific (2 mentions)
Benzonase, by Merck Group (2 mentions)
2 protocols using sds page 4 12 bis tris protein gels
1
Western Blot Analysis of Protein Lysates
2
Western Blot Analysis of Protein Lysates
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Cells were washed with PBS and lysed in RIPA lysis buffer (Thermo Fisher Scientific) with Halt Protease Inhibitor Cocktail (Thermo Fisher Scientific) and Benzonase (Sigma-Aldrich) for 20 min on ice. The insoluble fraction was removed by centrifugation; the protein concentration was quantified using a BCA protein assay kit (Thermo Fisher Scientific); and an equal amount of lysate was run on SDS-PAGE 4-12% Bis-Tris protein gels (Thermo Fisher Scientific) and then transferred to nitrocellulose membrane with an XCell II Blot Module Wet Tank Transfer System (Thermo Fisher Scientific). Membranes were blocked in Intercept (PBS) Blocking Buffer (LI-COR Biosciences) and incubated with primary antibodies overnight at 4 °C. The membranes were then washed in Tris-buffered saline with Tween 20 (TBS-T), incubated for 1 h with secondary IRDye-conjugated antibodies (LI-COR Biosciences) and washed three times in TBS-T for 5 min before near-infrared western blot detection on an Odyssey Imaging System with Image Studio software (LI-COR Biosciences).
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