L-Alanine Assay Kit, BioAssay(TM)

Alanine, a nonessential amino acid, is utilized in the glucose-alanine cycle between tissues and the liver. In tissues that metabolize amino acids, amino groups are collected as glutamate by transamination. The amine group is then transferred by alanine transaminase (ALT) from glutamate to pyruvate to form alanine and a-ketoglutarate. The alanine generated is transported to the liver where a reverse ALT reaction occurs and pyruvate is regenerated. Pyruvate is converted through gluconeogenesis to glucose which can then be recirculated to the tissues. Alanine concentration may have some correlation with high blood pressure, energy intake, cholesterol levels and body mass index.\n\nAlanine Assay Kit provides a simple, direct and automation-ready procedure for measuring alanine concentration. Alanine is converted into pyruvate which can then be directly measured. The color intensity of the reaction product at 570nm or fluorescence intensity at lem/ex=585/530nm is directly proportional to the alanine concentration in the sample.\n\nKey Features:\nSensitive and accurate. Linear detection range in 96-well plate: 1 to 200uM alanine for colorimetric assays and 0.4 to 20uM for fluorimetric assays.\n\nApplications:\nDirect Assays: alanine levels in plasma, serum, urine, tissue and culture media. Drug Discovery/Pharmacology: effects of drugs on alanine metabolism.\n\nKit Contents (100 Tests In 96-Well Plates):\nDeveloper: 6ml ALT Enzyme: 120ul\nDye Reagent: 120ul Cosubstrate: 600ul\nAlanine Standard: 400ul\nStorage conditions. The kit is shipped on ice. Store all kit components at -20 degrees C. Shelf life of six months after receipt.\nPrecautions: reagents are for research use only. Normal precautions for laboratory reagents should be exercised while using the reagents. Please refer to Material Safety Data Sheet for detailed information.\n\nProcedures:\nImportant:: equilibrate Developer to desired assay temperature. The assay requires 30 min when performed at 37 degrees C or 60 min if performed at RT (25 degrees C).\nSample Preparation\nTissue or cell samples (2

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SPECIFICATIONS

Catalog Number

A1205-13

Size

1Kit

References

1. Kiely, A. et al (2007). Pro-inflammatory cytokines increase glucose, alanine and triacylglycerol utilization but inhibit insulin secretion in a clonal pancreatic b-cell line. Journal of Endocrinology 195:113-23.\n2. Layman, DK (2003). The Role of Leucine in Weight Loss Diets and Glucose Homeostasis J. Nutr. 133: 261S