Z-Gly-Gly-Arg-AMC (acetate)

Z-Gly-Gly-Arg-AMC acetate is a thrombin-specific fluorogenic substrate for testing of thrombin generation in PRP and platelet-poor plasma (PPP) (Ex/Em = 390/480 nm)^[1][2]. In Vitro:1. Solution preparation^[3]
1.1 Preparation of stock solution
Solvent: H2O
Concentration: 10 mM.
Storage: Store at -20°C or -80°C in dark after aliquoting. Avoid repeated freezing and thawing.
1.2 Preparation of working solution
Dilute to 10 µM with PBS (or other buffer) (optimized according to the experiment).
Note: The working solution should be prepared and used immediately. Keep it away from light.

2. Thrombin inhibition screen
2.1 The samples and thrombin are transferred to a 384-well plate in sequence.
2.2 Add Z-Gly-Gly-Arg-AMC (50 μM) after incubation for 15 minutes.
2.3 Using a buffer containing 10 mM Tris-Cl (pH 7.4), 150 mM NaCl, 10 mM MgCl2, 1 mM CaCl2, 0.1% w/v BSA, 0.01% v/v Triton-X100, and 1% v/v DMSO and the fluorogenic substrate Z-Gly-Gly-Arg-AMC (50 μM).
2.4 The fluorescence intensity is measured with a fluorescence plate reader (Ex/Em = 360/465 nm) at 25°C for a period of 30 min with a read every 3 min.