Purified human thrombin, ATIII and factor Xa were purchased from Haematologic Technologies, Inc (Essex Junction, USA). Purified methylglyoxal was purchased from Sigma Aldrich (St. Louis, USA). Fluorogenic thrombin substrate, Z-Gly-Gly-Arg-AMC, was purchased from Bachem (Torrance, CA). Fluorogenic Xa substrate Boc-Ile-Glu-Gly-Arg-AMC was purchased from Bachem (Torrance, CA). Human standard plasma and ATIII-deficient plasma were provided by Affinity biological (Ancaster, CAN). PBS with a pH of 7.4 and 9 g/L sodium chloride, 0.795 g/L disodium phosphate and 0.144 g/L potassium dihydrogen phosphate was purchased from Corning (Midland, MI). Heparin with an average molecular weight of 4500 Da was purchased from Sanofi (Bridgewater, NJ).
Boc ile glu gly arg amc
Manufactured by Bachem
Sourced in Switzerland
Boc-Ile-Glu-Gly-Arg-AMC is a synthetic peptide substrate used for the analysis and detection of enzymatic activity. It consists of a sequence of five amino acids (isoleucine, glutamic acid, glycine, and arginine) with a tert-butyloxycarbonyl (Boc) protecting group at the N-terminus and 7-amino-4-methylcoumarin (AMC) at the C-terminus. This substrate can be used to measure the activity of various proteases that cleave the arginine-AMC bond, releasing the fluorescent AMC moiety.
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Lab products found in correlation
2 protocols using boc ile glu gly arg amc
1
Thrombin Inhibition Kinetics Assay
2
Fluorometric Assay for FXa Activity
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FXa activity was measured using a fluorometric assay based on the cleavage rate of the synthetic substrate Boc-Ile-Glu-Gly-Arg-AMC (50 μM, I-1100; Bachem, Bubendorf, Switzerland). Commercial bovine FXa (Haematologic Technologies, BCXA-1060) was diluted in buffer (in mM: 50 TRIS/HCl pH 8.0, 150 NaCl, 1 CaCl2, and 0.1% BSA) and placed in a 96 well black plate with and without FEAM (10 µM). The substrate (50 μM) was added to each well (the final volume in each well was 100 μl). The cleavage of the substrate was measured using a microplate reader at 37 °C, every 2 min for 25 cycles. The results are presented as a linear increase of fluorescence intensity over time, relative to control.
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