Experiments on biochemical analysis of cell death
Experiments on biochemical analysis of cell death
This experiment is based on the "Guide to Cellular Experiments", translated by Huang Peitang et al.
Operation method
Colorimetric quantification of caspase activation
Materials and Instruments
Cells Move 1. 250 g centrifugation for 5 minutes to collect 2X106~5X106 cells, wash once with cold PBS of pH 7.2, and resuspend with 200 μl of lysis buffer. For more product details, please visit Aladdin Scientific website.
PBS Eppendorf DMSO
Microtitre plates
2. Incubate cells on ice for 30 minutes.
3. centrifuge the samples at 13000 r/min for 5 min with Eppendorf and take the top sample for caspase activity assay: reserve one aliquot of each sample for protein quantification.
4. Dissolve the substrate in water or DMSO to make a 20 mmol/L storage solution that can be stored at -20°C protected from light.
5. Place the reaction mixture in a microtitre plate and read the reaction data on a SpectraMax microtitre plate analyzer with a kinetic device under a 405 nm filter. Determine the initial rate of hydrolysis of the substrate by testing the sample every 2-20 seconds; ensure that the resulting rate of hydrolysis of the substrate is linear.