| Assay Method Information | |
| | Biochemical Human MASP2 Assay |
| Description: | Recombinant human MASP2 enzyme produced in the HEK 293 cells was diluted in the reaction buffer (50 mM HEPES pH 8,0; 100 mM NaCl; 0,01% CHAPS; 0.5 mM Gluthathione) to the concentration of 20 nM and 25 μl was transferred into each single well of 384-well white microtiter plate (Greiner Bio One 781075). 1 μl of the inhibitor compound solution (dissolved in DMSO, at the corresponding concentration) or pure DMSO as a control was added to the same wells. The enzymatic reaction was initiated by addition of 25 μl of 60 μM solution of the FRET substrate DABCYL-KISPQGYGRR-Glu(EDANS) NH2; (Dabcyl-4-((4-(dimethylamino)phenyl)azo)benzoic acid; Edans-5-[(2-Aminoethyl) amino]naphthalene-1-sulfonyl; custom synthesis by Jerini Peptide Technologies, Berlin) in the reaction buffer. The microtiter plate was incubated for 60-120 min at the temperature of 32 C. |
| Affinity data for this assay | |
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