SH-SY5Y cells were seeded in 6-well plates at 10 6 cells/well for 24 hours and then treated with increasing MPP + concentrations for 24 hours. Cells were the gently washed with PBS and chemically detached with 0.05% trypsin/EDTA. After detachment, trypsin was quenched with complete media, cells pelleted by centrifugation and caspase activity detected with FAM-FLICA capase-1 assay kit (ImmunoChemistry Technologies, LLC, Bloomington, MN, USA) according to manufacturer's recommendations. In a nutshell, cells were stained were incubated with FLICA (1: 30) (link) for 1 hour at 37°C protected from light. Cells were then by washed with 5 volumes of 1X apoptosis wash buffer, pelleted by centrifugation, resuspended in 1X apoptosis wash buffer with 5µL of PI and incubated for 10 minutes 37°C protected from light. Data was acquired with Beckman DxFlex ow cytometer (Beckman, USA) and analyzed with CytExpert (Beckman Coulter Inc, CA, USA). Single stain controls were used to calculate compensation. PI positive cells were detected according to the operation procedure of PI staining kit (KeyGEN Biotech, NanJing,
Fam flica capase 1 assay kit
Manufactured by ImmunoChemistry Technologies
Sourced in United States
The FAM-FLICA caspase-1 assay kit is a laboratory product designed to detect and measure the activity of caspase-1, an enzyme involved in inflammatory processes. The kit utilizes a fluorescent probe that binds to active caspase-1, enabling the quantification of its presence in cell samples.
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2 protocols using fam flica capase 1 assay kit
1
Caspase-1 Activity Assay in MPP+ Treated SH-SY5Y Cells
2
Caspase-1 Activation in SH-SY5Y Cells
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SH-SY5Y cells were seeded in 6-well plates at 10 6 cells/well for 24 hours and then treated with increasing MPP + concentrations for 24 hours. Cells were the gently washed with PBS and chemically detached with 0.05% trypsin/EDTA. After detachment, trypsin was quenched with complete media, cells pelleted by centrifugation and caspase activity detected with FAM-FLICA capase-1 assay kit (ImmunoChemistry Technologies, LLC, Bloomington, MN, USA) according to manufacturer's recommendations. In a nutshell, cells were stained were incubated with FLICA (1:30) for 1 hour at 37°C protected from light. Cells were then by washed with 5 volumes of 1X apoptosis wash buffer, pelleted by centrifugation, resuspended in 1X apoptosis wash buffer with 5µL of PI and incubated for 10 minutes 37°C protected from light. Data was acquired with Beckman DxFlex ow cytometer (Beckman, USA) and analyzed with CytExpert (Beckman Coulter Inc, CA, USA). Single stain controls were used to calculate compensation. PI positive cells were detected according to the operation procedure of PI staining kit (KeyGEN Biotech, NanJing,
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