| ALX-850-274-KI01 | 1 Set | 1,081.00 USD |  |
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| Applications: | Colorimetric detection |
| Contents: | Contains the following components: - Caspase-1 Substrate, Ac-YVAD-pNA (125µl) - Caspase-2 Substrate, Ac-VDVAD-pNA (125µl) - Caspase-3 Substrate, Ac-DEVD-pNA (125µl) - Caspase-5 Substrate, Ac-WEHD-pNA (125µl) - Caspase-6 Substrate, Ac-VEID-pNA (125µl) - Caspase-8 Substrate, Ac-IETD-pNA (125µl) - Caspase-9 Substrate, Ac-LEHD-pNA (125µl) - Cell Lysis Buffer (100ml) - Dilution Buffer (160ml) - 2x Reaction Buffer (20ml) - DTT (1M) (400µl) |
| Quantity: | 7x25 tests |
| Handling: | For optimal storage temperature of single components please refer to individual label. Avoid freeze/thaw cycles. |
| Shipping: | Blue Ice |
| Short Term Storage: | -20°C |
| Protocol: | Assay Procedure: 1. Induce apoptosis in cells by desired method. Concurrently incubate a control culture without induction. 2. Count cells and pellet 1-5x106 cells. 3. Resuspend cells in 50 µl of chilled Cell Lysis Buffer and incubate cells on ice for 10 min. Centrifuge for 1 min in a microcentrifuge (10’000 x g). 4. Transfer supernatant to a fresh tube and assay protein concentration. 5. Dilute 100-300 µg protein to 50 µl Cell Lysis Buffer for each test. 6. Add 50 µl of 2x Reaction Buffer containing 10 mM DTT to each sample. 7. Add 5 µl of the 4 mM pNA conjugated substrates (200 µM final conc.) into each tube individually and incubate at 37°C for 1-2 hours. 8. Read samples at 400 or 405nm in a microtiter plate reader, or spectrophotometer using a 100 µl micro quartz cuvet (Sigma), or dilute sample to 1ml with Dilution Buffer and using regular cuvet (note: dilution of the samples proportionally decreases the reading). Fold-increase in caspase activity can be determined by comparing these results with the level of the uninduced control. Note: Background reading from cell lysates and buffers should be subtracted from the readings of both induced and the uninduced samples before calculating fold increase in caspase activity. |
| Regulatory Status: | RUO - Research Use Only |
| Compatibility: | This product is compatible with the Absorbance 96 Plate Reader. |
Absorbance 96 Plate Reader |
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| Personal, Compact Plate Reader | ||||||
| ELISA, Colorimetric detection | Print as PDF | ||||||
| ENZ-INS-A96 | 1 ea | Inquire for pricing | ||||
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