Liver tissues were homogenized in chilled lysis buffer (Thermo Fisher Scientific, Waltham, MA, USA), followed by incubation for 20 minutes on ice. After centrifugation at 12000 g for 10 minutes at 4°C, the supernatants were collected as protein samples. Protein content was determined using the Pierce™ BCA Protein Assay kit (Thermo Fisher Scientific, Waltham, MA, USA). Approximately 40 μg of protein was loaded into a 12% bis-tris polyacrylamide gel (Thermo Fisher Scientific, Waltham, MA, USA) for separation. Afterward, the proteins were transferred to a polyvinyl difluoride membrane. After blocking, the membranes were blotted with primary antibodies against caspase-1 (Abcam, San Francisco, CA, USA), which was followed by blotting with a secondary antibody in a standard fashion. The membranes were developed using enhanced chemical luminescence (Biological Industries, Kibbutz Beit-Haemek, Israel) and exposed to X-ray films. β-Actin served as a protein loading control.
Primary antibodies against caspase 1
Manufactured by Abcam
Sourced in United States
Primary antibodies against caspase-1. Caspase-1 is a protease enzyme involved in the inflammatory response and programmed cell death.
Automatically generated - may contain errors
Lab products found in correlation
Lysis buffer, by Thermo Fisher Scientific (1 mentions)
Enhanced chemical luminescence, by Sartorius (1 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (1 mentions)
Bis tris polyacrylamide gel, by Thermo Fisher Scientific (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Propidium iodide, by Merck Group (1 mentions)
2 protocols using primary antibodies against caspase 1
1
Quantification of Caspase-1 in Liver Tissue
2
Testicular Caspase-1 and NLRP3 Immunoreactivity
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The fixed testicular tissues were dehydrated in a graded series of ethanol and then the 5-μm thick sections were prepared from paraffin-embedded testicular tissues. After deparaffinization, the tissue sections were rehydrated in graded ethanol to determine caspase-1 and NLRP3 immunoreactivity. The tissue sections were permeabilized with 10 mM sodium citrate and 0.05% Tween 20, and blocked in a solution of 1% (w/v) BSA (Sigma-Aldrich, St. Louis, MO, USA) in PBS. The tissue sections were incubated overnight at 4 °C with primary antibodies against caspase-1 (1:1000 dilution, Abcam, Cambridge, MA) and NLRP3 (1:500 dilution, Abcam, Cambridge, MA). After the secondary antibody (1:500 dilution, Abcam, Cambridge, MA) incubation for 2 h at 37 °C, the cellular nuclei were stained with propidium iodide (PI; 1:1000, Sigma-Aldrich, St. Louis, MO, USA). The cell counting and merging pictures were done using “ImageJ” software (Image J, U. S. National Institutes of Health, Bethesda, Maryland, USA).
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