Alanine aminotransferase reagent kit GPT – 11 series

solutionfor clinical chemistrydiagnostic

For the quantitative determination of Alanine Aminotransferase in serum. METHODOLOGY The International Federation of Clinical Chemistry (IFCC) published a proposed recommended method in 1980 utilizing the LDH-NADH coupled assay. The procedure described herein is based on that method. Principle ALT catalyzes the transfer of the amino group from L-alanine to -ketoglutarate resulting in the formation of pyruvate and L-glutamate. Lactate dehydrogenase catalyzes the reduction of pyruvate and the simultaneous oxidation of NADH to NAD. The resulting rate of decrease in absorbance is directly proportional to ALT activity. REAGENT PREPARATION Prepare working reagent by mixing 4 parts of R1 reagent with 1 part R2 reagent. (e.g. 200 µL R1 with 50 µL R2 reagent.) REAGENT DETERIORATION Do not use reagent if: 1. The initial absorbance at 340nm is below 0,8. 2. The reagent fails to meet stated parameters of performance. PRECAUTIONS 1. This reagent set is for in vitro diagnostic use only. 2. The R1 reagent contains sodium azide as a preservative. Do not ingest. SPECIMEN COLLECTION AND STORAGE 1. Non-hemolyzed serum is recommended. Red cells contain ALT which can give falsely elevated results. 2. ALT in serum is stable for three days at room temperature (15-30°C), 1 week at 4°C. PROCEDURE Wavelength : 340 nm Assay Type : Kinetic Reaction Direction : Decreasing Temperature : 37°C Opthical path : 1 cm 1. Bring to room temperature ( 15 -30 °C) 2. Set the photometer to 0 (zero ) with distilled water 3. Mix and incubate 37 °C 4. After 60 sec. read and record the absorbance 5. Retun tube to 37 °C.