HepG2 cells in all groups were scraped in cell lysis buffer; for all groups, reaction buffer and YVAD-AFC substrate were further added to measure the activation of caspase-1 according to the instructions of a commercially available caspase-1 activity assay kit (Abcam, Cambridge, UK).
Caspase 1 activity assay kit
Manufactured by Abcam
Sourced in United Kingdom, United States
The Caspase-1 activity assay kit is a laboratory tool designed to measure the activity of the enzyme Caspase-1 in biological samples. Caspase-1 is a key mediator of the inflammatory response and plays a crucial role in various cellular processes. This kit provides a simple and efficient method to quantify Caspase-1 activity, allowing researchers to study its involvement in different biological systems and disease states.
Automatically generated - may contain errors
Lab products found in correlation
Triglyceride assay kit, by Nanjing Jiancheng (1 mentions)
Gsh assay kit, by Nanjing Jiancheng (1 mentions)
Total cholesterol assay kit, by Nanjing Jiancheng (1 mentions)
Dcfh da fluorescent probe kit, by Beyotime (1 mentions)
Mouse il 1β elisa kit, by R&D Systems (1 mentions)
Alexa fluor 488 annexin 5 dead cell apoptosis kit with alexa fluor 488 annexin 5 and pi for flow cytometry, by Thermo Fisher Scientific (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Bafilomycin a1, by Merck Group (1 mentions)
Ripa buffer, by Cell Signaling Technology (1 mentions)
Atg5 sirna, by Thermo Fisher Scientific (1 mentions)
Antibodies to cleaved caspase 3, by Cell Signaling Technology (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
Lipofectamine 2000 reagent, by Thermo Fisher Scientific (1 mentions)
Deadend colorimetric tunel assay kit, by Promega (1 mentions)
5 protocols using caspase 1 activity assay kit
1
Caspase-1 Activity Assay in HepG2 Cells
2
Caspase-1 Activity Assay in RAW264.7 Cells
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The caspase-1 activity assay was performed at first by scraping RAW264.7 cells in cell lysis buffer and then later adding reaction buffer and YVAD-AFC substrate by following the instructions provided by a commercially available caspase-1 activity assay kit (Abcam, Cambridge, UK).
3
Evaluation of Hepatocyte Oxidative Stress
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The cell culture supernatant of primary hepatocytes was collected for further analysis. Triglyceride (TG) and total cholesterol (T-CHO) were measured using a triglyceride assay kit and a total cholesterol assay kit, respectively (Nanjing Jiancheng Bioengineering Institute, China). The IL-1β concentration was measured using an IL-1β ELISA kit (Lianke Biotechnology Company, China). Plasma from mice was centrifuged, separated and stored at -80 °C for further analysis. Plasma levels of malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione (GSH) were measured using MDA, SOD and GSH assay kits, respectively (Nanjing Jiancheng Bioengineering Institute, China). The plasma level of reactive oxygen species (ROS) was measured using an ROS ELISA kit (Nanjing JiaBeiSen Biotechnology, China). ROS generation in the cell culture supernatant was assayed using a DCFH-DA fluorescent probe kit (Beyotime Biotechnology, China). Total protein was extracted from mouse liver tissues. Caspase-1 activity in liver tissues was assessed with a Caspase-1 activity assay kit (BioVision, Milpitas, CA, United States). All procedures were performed according to the manufacturers’ instructions.
4
Caspase-1 Activity and IL-1β Quantification
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Control and SEA-treated HSCs were washed twice with PBS, scraped in lysis buffer to extract proteins and measure caspase-1 activity according to manufacturer's instructions of a commercially available caspase-1 activity assay kit (Biovision). Cell supernatant was harvested and IL-1β level was measured by using a mouse IL-1β ELISA kit (R&D Systems) according to the manufacturer's instructions.
5
Apoptosis Mechanisms in Cell Cultures
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