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GGT Clinical Chemistry Reagent Enzymatic colorimetric method KINETIC PRINCIPLE Gamma-glutamyltransferase (?-GT) catalyzes the transfer of a ?- glutamyl group from ?-glutamyl-3-carboxy-4-nitroanilide to glycylglycine with the formation of L-?-glutamyl-glycylglycine and 5 amino-2-nitro-benzoate. The amount of 5-amino-2-nitro-benzoate formed, monitored kinetically at 405 nm, is proportional to the enzyme activity present in the sample. REAGENT COMPOSITION 4 x 100 ml R1 Buffer/Glycylglycine. TRIS 133 mmol/L pH 8.2, glycylglycine 138 mmol/L, sodium azide 0.09%. R2 Substrate/Glupa-C. L-?-glutamyl-3-carboxy-4-nitroanilide 23 mmol/L, ethylene glycol 900 mmol/L, sodium azide 0.09%. STORAGE AND STABILITY Store at 2-8?C. REAGENT PREPARATION Working reagent. Mix 4 mL of R1 + 1 mL of R2. Stable for 3 weeks at 2-8?C or for 5 days at 15-25?C. Protect from light. SAMPLES Serum or EDTA plasma free of hemolysis. Fluoride, citrate and oxalate inhibit ?-GT activity2 . The enzyme in the sample is stable for at least 1 week at 2-8?C and for at least 2 months when frozen. Only freeze once. Discard contaminated specimens. The samples should be handled as potentially infectious in accordance with Good Manufacturing Practices. MATERIALS REQUIRED ? Photometer or spectrophotometer with cuvette thermostatic compartment set at 25/30/37?C, capable of reading at 405 nm. ? Stopwatch, strip-chart recorder or printer. ? Cuvettes with 1-cm pathlength. ? Pipettes to measure reagent and samples. ? Ref. 1975005 Human Multicalibrator (optional). ? General laboratory equipment. GGT Clinical Chemistry Reagent per? box