Isopeptidase-Fluorogenic
Assays

Art. No.

F001

Synonym

ZediXcite 330/418 Fluorogenic FXIII-Assay Kit

Description

Isopeptidase-activity based ready to use kit for fluorescent measurement optimized for Factor XIII (FXIII) in plasma samples. FXIIIa cleaves a dark quenching molecule (2,4-dinitrophenyl) from the side chain of a modified peptide incorporating glycine methyl ester. Subsequently, the fluorescence of an N-terminal coupled fluorophore (2-amino benzoic acid, 2-Abz) increases and can be monitored online (excitation wavelength 313-330 nm; emission wavelength 418 nm). The assay is suitable for research purposes, drug discovery programs, and quality assurance.

Assay principle

Factor XIII is converted by Thrombin into Factor XIIIa. At the same time, Thrombin converts Fibrinogen into Fibrin. The clotting is prevented by an aggregation inhibiting peptide. Factor XIIIa cleaves a dark quenching molecule from the side chain of a modified peptide incorporating glycine methyl ester. Subsequently, the fluorescence of an N-terminal coupled dye increases and can be monitored on-line (excitation wavelength 313 nm; emission wavelength 418 nm). Basically, the isopeptidase activity of Factor XIIIa was described by Parameswaran et al., the modified peptide used is intellectual property of Zedira.