Cells were lysed in RIPA lysis buffer with 2 mM phenylmethylsulfonyl fluoride, 1 mM sodium orthovanadate and protease inhibitor cocktail (Santa Cruz Biotech, Dallas, TX, sc-24948) for 30 min at 4 °C and then centrifuged at 16,000 g for 15 min. Samples (25–50 μg of protein/lane) were resolved by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and transferred to nitrocellulose membranes (Bio-Rad, Hercules, CA, 1704271). Blots were probed with rabbit monoclonal anti-caspase 1 (1:1000; Abcam, Cambridge, UK, ab179515), rabbit polyclonal anti-IL-1beta/IL-1F2 (1:1000; Novus Biologicals, Abingdon, UK, NB600-633), rabbit polyclonal anti-NLRP1/NALP1 (1:1000; Cell Signaling, Danvers, MA, 4990S), rabbit polyclonal anti-NLRP3/NALP3 (1:2500; Novus Biologicals, NBP2-12446), and mouse monoclonal anti-ACTB/actin (1:30,000; Sigma-Aldrich, A3853). After overnight incubation at 4 °C, bound antibodies were visualized using horseradish peroxidase-coupled secondary antibodies and the Clarity™ Western ECL Substrate (Bio-Rad, 1705061) or Clarity™ Max ECL western blotting substrate (Bio-Rad, 1705062) for low protein concentrations.
Anti nlrp1 nalp1
Manufactured by Cell Signaling Technology
Anti-NLRP1/NALP1 is a primary antibody that recognizes the NLRP1/NALP1 protein. NLRP1/NALP1 is a NOD-like receptor that plays a role in the regulation of the inflammasome complex.
Automatically generated - may contain errors
Lab products found in correlation
Protease inhibitor cocktail, by Santa Cruz Biotechnology (2 mentions)
Clarity max western ecl blotting substrate, by Bio-Rad (2 mentions)
Anti il 1beta il 1f2, by Novus Biologicals (2 mentions)
Nbp2 12446, by Merck Group (2 mentions)
Nitrocellulose membrane, by Bio-Rad (2 mentions)
Clarity western ecl substrate, by Bio-Rad (2 mentions)
2 protocols using anti nlrp1 nalp1
1
Western Blot Analysis of Inflammasome Proteins
2
Western Blot Analysis of Inflammasome Proteins
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Cells were lysed in RIPA lysis buffer with 2 mM phenylmethylsulfonyl fluoride, 1 mM sodium orthovanadate and protease inhibitor cocktail (Santa Cruz Biotech, Dallas, TX, sc-24948) for 30 min at 4 °C and then centrifuged at 16,000 g for 15 min. Samples (25–50 μg of protein/lane) were resolved by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and transferred to nitrocellulose membranes (Bio-Rad, Hercules, CA, 1704271). Blots were probed with rabbit monoclonal anti-caspase 1 (1:1000; Abcam, Cambridge, UK, ab179515), rabbit polyclonal anti-IL-1beta/IL-1F2 (1:1000; Novus Biologicals, Abingdon, UK, NB600-633), rabbit polyclonal anti-NLRP1/NALP1 (1:1000; Cell Signaling, Danvers, MA, 4990S), rabbit polyclonal anti-NLRP3/NALP3 (1:2500; Novus Biologicals, NBP2-12446), and mouse monoclonal anti-ACTB/actin (1:30,000; Sigma-Aldrich, A3853). After overnight incubation at 4 °C, bound antibodies were visualized using horseradish peroxidase-coupled secondary antibodies and the Clarity™ Western ECL Substrate (Bio-Rad, 1705061) or Clarity™ Max ECL western blotting substrate (Bio-Rad, 1705062) for low protein concentrations.
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