Ac-WEHD-AFC TFA is a fluorogenic caspase-1 fluorogenic activity and can be used for the research of tumor and inflammation[1].
Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
Caspase-1 Fluorogenic Activity Assay[1]:
1. Incubate the cells according to your normal protocol.
2. Cells are transiently transfected with caspase-1 in combination with pcDNA3.1, Myc-cIAP2, Myc-cIAP1, or Myc-XIAP and collected 24 hr later.
3. Cells were lysed in CHEGG buffer before sonicating for 15 s at 60% amplitude (for cleavage assays).
4. Lysates were incubated with 10 mM of the fluorogenic caspase-1 substrate Ac-WEHD-AFC TFA.
5. The release of free AFC was monitored continuously for 1 hr (excitation 380 nm, emission 460 nm) in 1 min intervals and expressed as arbitrary fluorescence units per minute.
[1]. Katherine LabbÉ, et al. Cellular inhibitors of apoptosis proteins cIAP1 and cIAP2 are required for efficient caspase-1 activation by the inflammasome. Immunity. 2011 Dec 23;35(6):897-907.