Transglutaminases are a family of enzymes that catalyse the posttranslational modification of proteins by inserting an isopeptide bond within or between polypeptide chains (Folk, J. E. and Cole, P. W., 1966). These enzymes catalyse the acyl transfer reaction between the γ-carboxyamide group of peptide-bound glutamine residues and a variety of primary amines, particularly the ε-amino group of lysine (Lorand L. et al., 1962). The resulting cross link is of great significance, since it is highly stable and also resistant to mechanical and proteolytic degradation.
The MICROBIAL TRANSGLUTAMINASE ASSAY KIT uses Z-Gln-Gly as the amine acceptor substrate and hydroxylamine as amine donor. In the presence of MTG hydoxylamine is incorporated into Z-Gln-Gly to form Z-glutamylhxdroxamate-glycine which develops a colored complex with iron (III) detectable at 525 nm.
(A1) ACTIVITY REAGENT 1: lyophilized TRIS buffer containing Z-Gln-Gly and reduced glutathione, 3 vials
(A2) ACTIVITY REAGENT 2: Hydroxylamine, 3 vials
(S) STOP REAGENT: Hydrochloric acid [4% v/v], Iron (III) chloride, 3 vials
(P1) MICROBIAL TRANSGLUTAMINASE (positive control): 3 vials
The MICROBIAL TRANSGLUTAMINASE ASSAY KIT can be used in standard spectrophotometers with polystyrene 1 mL cuvettes. Refer to the instructions of the manufacturer.
P1 has to be stored at -20°C (shipment is possible at 4 – 8°C).
A1, A2, S has to be stored at 4 – 8°C.
The unopened reagents are stable until the expiration date printed on the box.