Caspase 3 Assay Kit, Colorimetric
SIGMA/CASP3C
Synonym: Colorimetric Caspase-3 Activity Assay Kit
Product Type: Chemical
Caspase 3 activity in apoptotic Jurkat cells. Apoptosis was induced in Jurkat human cells with 1 μg/ml staurosporine for 0, 1, 2, and 3 hours. The cells were lysed in 1X lysis buffer at a ratio of 100 μl per 107 cells. The caspase 3 activity in the lysate was determined using the Caspase 3 Colorimetric Assay Kit (CASP-3-C). Ten μl of each lysate was tested both with and without caspase 3 inhibitor, in a total reaction volume of 100 μl using 96 well multiwell plates. The substrate (Ac-DEVD-pNA) concentration was 200 μM, and the inhibitor (Ac-DEVD-CHO) concentration was 0.05 μM. The assay was performed at pH 7.4 at 37 °C for 42 minutes.
| Quality Level | 300  |
| shipped in | dry ice |
| storage temp. | −20°C |
| usage | sufficient for 1,000 multiwell tests (in 96-well multiwell plates) |
| sufficient for 100 standard tests (1 ml) |
| Application: | The assay can be performed (a) in 1 mL volume and measured using a spectrophotometer, or (b) in 100 µL volume in a 96-well plate using an ELISA reader. |
| General description: | Caspase 3 Colorimetric Assay Kit provides the reagents needed for a quick and efficient detection of caspase 3 activity in cell lysates and in purified preparations of caspase 3. The Caspase 3 Colorimetric Assay Kit is based on the hydrolysis of acetyl-Asp-Glu-Val-Asp p-nitroanilide (Ac-DEVD-pNA) by caspase 3, resulting in the release of the p-nitroaniline (pNA) moiety. p-Nitroaniline is detected at 405 nm (εmM=10.5). The concentration of the pNA released from the substrate is calculated from either the absorbance values at 405 nm or from a calibration curve prepared with pNA standards (pNA standard included with the kit). |
| Other Notes: | One unit of caspase 3 will cleave 1.0 μmol of Ac-DEVD-pNA per min at pH 7.4 at 25 °C. |
| Other Notes: | Use 17 megohm water only |
| Symbol |   GHS06,GHS08 |
| Signal word | Danger |
| Hazard statements | H301 + H311 + H331 - H373 - H412 |
| Precautionary statements | P260 - P273 - P280 - P301 + P310 + P330 - P302 + P352 + P312 - P304 + P340 + P311 |
| Hazard Codes | T |
| Risk Statements | 23/24/25-33-52/53 |
| Safety Statements | 36/37-45-61 |
| RIDADR | UN 3316 9 |
| Flash Point(F) | 415.4 °F - closed cup |
| Flash Point(C) | 213 °C - closed cup |
| Storage Temp. | −20°C |
| UNSPSC | 12161503 |
| Components | Ac-DEVD-CHO inhibitor .5 mg; Ac-DEVD-pNA substrate 15 mg; p-Nitroaniline standard 1 mg; Water (17-18 megaOhm) 125 mL |