Ac-LEHD-AFC 

Ac-LEHD-AFC is a fluorescent substrate for caspase-9, can be used to assess the activity of caspase^[1].

Caspase-9^[1]

Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
The assessment of the caspase activity by fluorogenic substrate assay^[1]:
1. Preparation of cell lysis buffer: 1 % Nonidet P-40, NaCl 200 mM, Tris/HCl 20 mM, pH 7.4, Leupeptin 10 mg/L, Aprotinin (HY-P0017) (0.27 kU/L) and 100 mM PMSF (HY-B0496) .
2. Incubate the lysis buffer (25 mg of total protein) with 50 mmol/L of AC-DEVD-AMC (HY-P1003) (caspase-3), AC-VEID-AFC (caspase-8) or AC-LEHD-AFC (caspase-9) in the buffer (HEPES 10 mM, pH 7.4, containing Mannitol 220 mM, Sucrose 68 mM, NaCl 2 mM, KH₂PO₄ 2.5 mM, EGTA 0.5 mM, MgCl₂ 2 mM, Pyruvate 5 mM, PMSF 0.1 mM, and Dithiothreitol 1 mM).
3. Assess the caspase activity by measuring fluorescent 7-amino-4-methylcoumarin released for 1 h at a 2-min intervals by a spectrofluorometer.

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

765.69

C₃₃H₃₈F₃N₇O₁₁

210345-03-2

454

445

FC(C(C1=CC=C(C=C1O2)NC([C@@H](NC([C@H](CC3=CNC=N3)NC([C@@H](NC([C@H](CC(C)C)NC(C)=O)=O)CCC(O)=O)=O)=O)CC(O)=O)=O)=CC2=O)(F)F

Room temperature in continental US; may vary elsewhere.

Please store the product under the recommended conditions in the Certificate of Analysis.

• [1]. Chae HJ, et al. Je-chun-jun induced apoptosis of human cervical carcinoma HeLa cells. Acta Pharmacol Sin. 2004 Oct;25(10):1372-9.  [Content Brief]