Apoptosis. Apoptosis was induced by incubating Jurkat cells with 0.2 µg/ml of anti-Fas monoclonal antibody for 8 hours. The Caspase 3 colorimetric assay was performed.

描述

The Caspase-3 colorimetric protease assay provides a simple and convenient means for quantitating caspases that recognize the amino acid sequence, DEVD. The kit includes substrate and optimized buffers. The substrate, DEVD-pNA, is composed of the chromophore p-nitroanilide (pNA), and a synthetic tetrapeptide, DEVD (Asp-Glue-Val-Asp), which is the upstream amino acid sequence of the Caspase-3 cleavage site in PARP. Upon cleavage of the substrate by Caspase-3 or related caspases, free pNA light absorbance can be quantified using a spectrophotometer or a microplate reader at 400 or 405 nm. Comparison of the absorbance of pNA from apoptotic sample with an uninduced control allows determination of the fold increase in Caspase-3 activity.

内容和储存

Kit contains:
• Cell Lysis Buffer (25 mL): Tris buffered saline (TBS) -containing detergent
• 2X Reaction Buffer (2 mL): contains buffered saline, glycerol and detergent
• Substrate (125 µL): contains 4 mM of synthetic peptide (sequence: DEVD) conjugated to the chromophore pNA in DMSO
• Dilution Buffer (25 mL): buffered solution containing HEPES, Tris and NaCl
• DTT (100 µL): contains 1 M dithiothreitol

Store kit at -20°C. Once opened, store Cell Lysis Buffer, 2X Reaction Buffer, and Dilution Buffer at 2 to 8°C. Refer to reagent label for expiration.

详细说明

常用规格