包装 | 价格(元) |
1mg | 电议 |
5mg | 电议 |
10mg | 电议 |
Cell experiment: | For the detection of caspase-3 activity, PBS washes cell pellets (derive from either the medium or the adherent cells) which are suspended in extract buffer [25 mM HEPES (pH7.4), 0.1% TritonX-l00, 10% glycerol, 5 mM DTT, 1mM phenylmethylsulfonyl fluoride, 10 mg/mL pepstatin, and 10 mg/mL Leupeptin] and vortexed vigorously. 20μl of extract (corresponding to 10% of the sample) are incubated with the caspase-3 fluorogenic substrates Ac-DEVD-AFC at 100 μM final concentration at room temperature, and caspase-3 activity is measured continuously by monitoring the release of fluorigenic AFC at 37℃[1]. |
产品描述 | Ac-DEVD-AFC is a fluorogenic substrate for activated caspase-3 (CPP32) with Km value of 9.7 μM, and related cysteine proteases [1][2]. Apoptosis is a process of programmed cell death that occurs in multicellular organisms. Caspase are a family of protease enzymes playing essential roles in programmed cell death (including apoptosis, pyroptosis and necroptosis) and inflammation. Caspase activation is a major event in apoptosis. Caspase-3 cleaves and activates caspases 6 and 7, and is processed and activated by caspases 8, 9, and 10 [1][2][3]. Ac-DEVD-AFC (N-Acetyl-Asp-Glu-Val-Asp-7-amido-4-Trifluoromethylcoumarin) is a fluorogenic substrate for activated caspase-3 and related caspases. During apoptosis, activated caspase-3 cleaves poly (ADP-ribose) polymerase, which it specifically targets at the amino sequence Asp-Glu-Val-Asp (DEVD). Sequence is based on PARP cleavage at Asp216 for caspase-3 [1][2]. Caspase activity can be quantified by fluorescent detection of free AFC (7-amino-4-trifluoromethylcoumarin) at excitation 400 nm and emission 505 nm [2]. References: |