Caspase-9 Fluorometric Assay Kit
Cysteine-containing aspartate proteolytic enzymes (Caspase) are a family of cysteine proteases that play important roles in apoptosis, necrosis, and inflammation. Caspase-9 is a priming type of caspase that initiates endogenous apoptosis, which is apoptosis of the mitochondrial pathway. When cells are stressed, Cytochrome c is released into the cytoplasm and binds to the apoptotic protein activator 1 (Apaf-1) to form a complex. Caspase-9 was then recruited into this complex, forming the apoptotic bodies. In apoptotic bodies, caspase-9 is activated by self-cleavage. Activated caspase-9 further activates downstream effector caspases, such as caspase-3 and caspase-7.
Caspase-9 Fluorometric Assay Kit provides a convenient and simple way for detecting the LEHD-dependent caspase activity. LEHD-AFC (AFC:7-amino-4-trifluoromethyl coumarin) emits blue light (λmax = 400 nm); while cleavage of LEHD-AFC by Caspase-9 or related caspases, free AFC emits a yellow-green fluorescence (λmax = 505 nm), which can be quantified by using a microplate reader or a fluorometer. Comparison of the fluorescence of AFC from an apoptotic sample with an uninduced control determines the fold increase in Caspase-9 activity.
| Components |
K2018-25T |
K2018-100T |
K2018-200T |
K2018-400T |
|
Cell Lysis Buffer |
25 mL |
100 mL |
100 mL |
100 mL |
|
2X Reaction Buffer |
2 mL |
4 X 2 mL |
16 mL |
32 mL |
|
LEHD-AFC (1 mM) |
125 µL |
500 µL |
2X 0.5 mL |
2X 1 mL |
|
DTT (1 M) |
100 µL |
400 µL |
400 µL |
400 µL |
|
Store the kit at -20°C, stable for 6 months. LEHD-AFC (1 mM) should be stored away from light. |
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| Features |
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