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).
Xcell 2 blot module wet tank transfer system
Manufactured by Thermo Fisher Scientific
The XCell II Blot Module Wet Tank Transfer System is a lab equipment product designed for protein transfer from electrophoresis gels to membranes. It provides a controlled environment for efficient protein transfer during Western blotting procedures.
Automatically generated - may contain errors
Lab products found in correlation
Intercept pbs blocking buffer, by LI COR (2 mentions)
Ripa lysis buffer, by Thermo Fisher Scientific (2 mentions)
Odyssey imaging system, by LI COR (2 mentions)
Irdye conjugated antibody, by LI COR (2 mentions)
Benzonase, by Merck Group (2 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Halt protease inhibitor cocktail, by Thermo Fisher Scientific (2 mentions)
Sds page 4 12 bis tris protein gels, by Thermo Fisher Scientific (2 mentions)
2 protocols using xcell 2 blot module wet tank transfer system
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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