Activated Caspase-1 was detected with the Pyroptosis/Caspase-1 Assay Kit (#9158) from ImmunoChemistry Technologies (CA, USA) according to the manufacturer’s instructions. CD4+ T cells were incubated with FLICA solution at 37 °C for 1 h. CD4+ T cells were then incubated with FITC anti-human CD4 antibody (357,406) from BioLegend (CA, USA) for 30 min. Fluorescently labeled cells were immediately detected by flow cytometry (Agilent, Novocyte 2060R).
Pyroptosis caspase 1 assay kit
Manufactured by ImmunoChemistry Technologies
Sourced in United States
The Pyroptosis/Caspase-1 Assay Kit is a laboratory tool designed to measure the activity of caspase-1, an enzyme involved in the process of pyroptosis, a form of programmed cell death. The kit provides a quantitative assessment of caspase-1 activity in cell lysates or other biological samples.
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3 protocols using pyroptosis caspase 1 assay kit
1
Detecting Activated Caspase-1 in CD4+ T Cells
2
Quantifying Pyroptosis via Caspase-1 Activity
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FLICA assay was performed using Pyroptosis/Caspase-1 Assay Kit (ImmunoChemistry Technologies) according to the manufacturer’s protocols. Briefly, THP-1 cells transfected with mock or β2MG expression vector were incubated with a green fluorescent carboxyfluorescein dye-labeled caspase-1 inhibitor (FAM-YVAD-FMK) for 1 h at 37 °C and washed with cellular wash buffer for three times. Then samples were suspended in cellular wash buffer, and fluorescence intensity was measured on a microplate reader (Powerscan®HT; DS Pharma Biomedical) set at 485 nm excitation and 528 nm emission.
3
Quantifying Pyroptosis via Caspase-1 Assay
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Cells with active caspase-1 were identified using the pyroptosis/caspase-1 assay kit (Immunochemistry Technologies, Bloomington, MN, USA) according to the manufacturer’s instruction. Shortly, the H9c2 cells were trypsinized to create suspensions, and then incubated with FLICA (FAM-YVAD-FMK) probe for 1 h at 37 °C. After three washes with washing buffer, the cells were resuspended in 200 μl PBS and analyzed immediately using a FACS Aria flow cytometer (BD Biosciences, San Jose, CA, USA).
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