Lysates were prepared from cell pellets in RIPA lysis buffer (Sigma-Aldrich, R0278). Protein determination was performed with Pierce BCA Protein Assay Kit (Thermo Fisher Scientific, 23227). Proteins from lysates were separated on Bis–Tris 12.5% acrylamide gels. Gels were transferred to nitrocellulose membranes (BioRad) and probed with rabbit anti-mouse Caspase-1 antibody (Santa Cruz, sc-514, 1 µg/ml, diluted 1:200) in 4 °C overnight, followed by incubation with HRP-conjugated secondary antibody (Jackson ImmunoResearch Laboratories, 111-035-144/062, 80 ng/ml) for 2 h at room temperature. Following membrane stripping, membranes were incubated with anti-mouse Gasdermin D (Abcam, ab209845, diluted 1:500) or with anti-mouse β-actin (Cell Signalling CST#4967, diluted 1:2000).
Rabbit anti mouse caspase 1 antibody
Manufactured by Santa Cruz Biotechnology
The Rabbit anti-mouse Caspase-1 antibody is a primary antibody that detects the Caspase-1 protein in mouse samples. Caspase-1 is a protease enzyme involved in the inflammatory response and programmed cell death processes.
Automatically generated - may contain errors
Lab products found in correlation
Hrp conjugated secondary antibody, by Jackson ImmunoResearch (1 mentions)
Nitrocellulose membrane, by Bio-Rad (1 mentions)
Ripa lysis buffer, by Merck Group (1 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (1 mentions)
Supersignal west pico kit, by Thermo Fisher Scientific (1 mentions)
Goat anti rabbit hrp, by Jackson ImmunoResearch (1 mentions)
Nupage gel, by Thermo Fisher Scientific (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
2 protocols using rabbit anti mouse caspase 1 antibody
1
Caspase-1 and Gasdermin D Detection
2
Caspase-1 Activation in Infected BMDMs
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BMDMs were seeded into 24-well plates at a density of 3 × 105 cells per well and infected with bacteria as described above. 3 h after infection, cells were lysed in 20 mM Hepes, 150 mM NaCl, 10% glycerol, 1% Triton X-100, and 1 mM EDTA. Lysates were mixed with protein loading buffer, boiled, centrifuged, and 20% of the total cell lysate loaded onto 4–12% NuPAGE gels (Invitrogen). Proteins were transferred to PVDF membrane (Millipore) and blotted with rabbit anti–mouse caspase-1 antibody (Santa Cruz Biotechnology, Inc.). Secondary antibody was goat anti–rabbit HRP (Jackson ImmunoResearch Laboratories, Inc.). The membrane was exposed on radiographical film using SuperSignal West Pico kit (Thermo Fisher Scientific).
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