Activity Assay Kits

MTG-ANiTA-KIT


(Ammonium-NicotinamidADPH-GLDH-Transglutaminase-Assay)

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1 Kit 575 € available


Documents
Art. No. M001
Background info 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.

MTG activity assay ANITA
Assay principle The MTG-ANiTA-KIT uses β-Casein as high molecular weight substrate. Transglutaminase catalyzes the formation of isopeptide bonds between protein-bound glutamine and lysine releasing one ammonium molecule per isopeptide bond formed.
In the indicator reaction the amount of released ammonium produced by transglutaminase is monitored in a glutamate dehydrogenase (GLDH) catalysed NADPH-dependent reaction. The consumption of NADPH is measured spectrophotometrically by the decrease of absorbance at 340 nm.
Reagents in the kit BD-BUFFER, 2 vials
[GLDH, ADP, α-KETOGLUTARATE]-MIX, 2 vials
NADPH (AD-REAGENT), 2 vials
β-Casein (TR-BUFFER), 2 vials
MTG-BLOCKER (INHIBITOR REAGENT), 2 vials
Equipment The MTG-ANiTA-KIT can be used in standard spectrophotometers with UV-cuvettes (e.g. BRAND Cat. No.:7591 50, semi-micro).
Refer to the instructions of the manufacturer.
Intended use Determination of microbial transglutaminase activity (MTG).
Kit is sufficient for 2 x 11 measurements.
Storage MTG-ANiTA-KIT should be stored at 4 – 8°C. The unopened reagents are stable until the expiration date printed on the box.
Reference(s) Lorand L., Konishi K., Jacobsen A., Nature 1962;194:1148-1149.
Folk, J. E. und Cole, P. W., Biochim. Biophys. Acta 1966, 122, 244-64.
Note INTENDED FOR RESEARCH USE ONLY, NOT FOR USE IN HUMAN, THERAPEUTIC OR DIAGNOSTIC APPLICATIONS.

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