Z-Gly-Gly-Arg-AMC 

Z-Gly-Gly-Arg-AMC 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].

Guide (The following is our recommended protocol. This protocol is only a guide and should be modified according to your specific needs).
1. Solution preparation^[3]
1.1 Preparation of stock solution
Solvent: DMSO
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.

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

579.60

C₂₈H₃₃N₇O₇

480

390

66216-78-2

Solid

White to off-white

Z-Gly-Gly-{Arg-AMC}

ZGG-{Arg-AMC}

Room temperature in continental US; may vary elsewhere.

polypeptide, sealed storage, away from moisture and light

*In solvent : -80°C, 6 months; -20°C, 1 month (polypeptide, sealed storage, away from moisture and light)

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

• [1]. Tarandovskiy ID, et al. Antiplatelet agents can promote two-peaked thrombin generation in platelet rich plasma: mechanism and possible applications. PLoS One. 2013;8(2):e55688.  [Content Brief]

  [2]. Machlus KR, et al. Effects of tissue factor, thrombomodulin and elevated clotting factor levels on thrombin generation in the calibrated automated thrombogram. Thromb Haemost. 2009 Nov;102(5):936-44.  [Content Brief]

  [3]. Mothukuri GK, et al. Macrocycle synthesis strategy based on step-wise "adding and reacting" three components enables screening of large combinatorial libraries. Chem Sci. 2020 Jun 26;11(30):7858-7863.  [Content Brief]