Z-Gly-Pro-AMC

• CAT Number: I043055
• CAS Number: 68542-93-8
• Molecular Formula: C25H25N3O6
• Molecular Weight: 463.48
• Purity: ≥95%
• Synonyms: benzyl N-[2-[(2S)-2-[(4-methyl-2-oxochromen-7-yl)carbamoyl]pyrrolidin-1-yl]-2-oxoethyl]carbamate
• InChI: InChI=1S/C25H25N3O6/c1-16-12-23(30)34-21-13-18(9-10-19(16)21)27-24(31)20-8-5-11-28(20)22(29)14-26-25(32)33-15-17-6-3-2-4-7-17/h2-4,6-7,9-10,12-13,20H,5,8,11,14-15H2,1H3,(H,26,32)(H,27,31)/t20-/m0/s1
• InChIKey: YWOXKKRKNGWXEG-FQEVSTJZSA-N
• SMILES: CC1=CC(=O)OC2=C1C=CC(=C2)NC(=O)C3CCCN3C(=O)CNC(=O)OCC4=CC=CC=C4

Z-Gly-Pro-AMC is a fluorogenic substrate. Z-Gly-Pro-AMC is hydrolyzed by prolyl endopeptidase to generate highly fluorescent 7-amido-4-methylcoumarin (HY-D0027). (λex=380 nm, λem=465 nm)[1].
Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
Z-Gly-Pro-AMC assay[1]:
1. Make a 5 μL of plasma sample is pre-incubated with 10 μL of 250 nM FAP inhibitor, 10 μL of 250 nM PREP inhibitor or 10 μL of 0.0025% (v/v) DMSO for 15 min at 37 ℃.
2. Add 35 μL pre-heated Z-Gly-Pro-AMC (380 μM diluted in buffer) to obtain a final concentration of 266 μM.
3. Fluorescence is measured kinetically for 30 min at 37 ℃.

Reference

[1]. Bracke A, et, al. The development and validation of a combined kinetic fluorometric activity assay for fibroblast activation protein alpha and prolyl oligopeptidase in plasma. Clin Chim Acta. 2019 Aug;495:154-160.
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