Caspase-4 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-1/4/5/11 are very important in inflammatory response and pyroptosis. Caspase-4/5 can recognize and bind intracellular lipopolysaccharide (LPS), which subsequently activate inflammasomes, which in turn promote the maturation and release of inflammatory mediator IL-1β, leading to an inflammatory response. In addition, activated caspase-4/5 can also cleave the substrate protein GSDMD, triggering the formation of cell membrane pores, resulting in the release of cell contents, and ultimately pyroptosis.
Caspase-4 Fluorometric Assay Kit provides a convenient and simple way for detecting the LEVD-dependent caspase activity. LEVD-AFC (AFC:7-amino-4-trifluoromethyl coumarin) emits blue light (λmax = 400 nm); while cleavage of LEVD-AFC by Caspase-4 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-4 activity.
| Components |
K2198-25T |
K2198-100T |
K2198-200T |
K2198-400T |
|
Cell Lysis Buffer |
25 mL |
100 mL |
100 mL |
100 mL |
|
2X Reaction Buffer |
2 mL |
4 X 2 mL |
16 mL |
32 mL |
|
LEVD-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. LEVD-AFC (1 mM) should be stored away from light. |
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| Features |
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