Ac-WEHD-AFC TFA

• CAT Number: I042048
• Molecular Formula: C40H38F6N8O13
• Molecular Weight: 952.77
• Purity: ≥95%
• Synonyms: (4S)-4-[[(2S)-2-acetamido-3-(1H-indol-3-yl)propanoyl]amino]-5-[[(2S)-1-[[(2S)-3-carboxy-1-oxo-1-[[2-oxo-4-(trifluoromethyl)chromen-7-yl]amino]propan-2-yl]amino]-3-(1H-imidazol-5-yl)-1-oxopropan-2-yl]amino]-5-oxopentanoic acid;2,2,2-trifluoroacetic acid
• InChI: InChI=1S/C38H37F3N8O11.C2HF3O2/c1-18(50)45-27(10-19-15-43-25-5-3-2-4-22(19)25)36(58)47-26(8-9-31(51)52)34(56)48-28(11-21-16-42-17-44-21)37(59)49-29(14-32(53)54)35(57)46-20-6-7-23-24(38(39,40)41)13-33(55)60-30(23)12-20;3-2(4,5)1(6)7/h2-7,12-13,15-17,26-29,43H,8-11,14H2,1H3,(H,42,44)(H,45,50)(H,46,57)(H,47,58)(H,48,56)(H,49,59)(H,51,52)(H,53,54);(H,6,7)/t26-,27-,28-,29-;/m0./s1
• InChIKey: UELPUHYAUOLMOR-MROKUPSKSA-N
• SMILES: CC(=O)NC(CC1=CNC2=CC=CC=C21)C(=O)NC(CCC(=O)O)C(=O)NC(CC3=CN=CN3)C(=O)NC(CC(=O)O)C(=O)NC4=CC5=C(C=C4)C(=CC(=O)O5)C(F)(F)F.C(=O)(C(F)(F)F)O

Ac-WEHD-AFC TFA is a fluorogenic caspase-1 substrate. Ac-WEHD-AFC TFA can measure 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.

Reference

[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.
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