Glyoxalase I Assay Kit, BioAssay(TM)

Glyoxalase I (GLO-1), a lactoylglutathione lyase also known as methylglyoxalase, aldoketomutase, ketone-aldehyde mutase, and (R)-Slactoylglutathione methylglyoxal-lyase, is an enzyme that catalyzes the isomerization of hemithioacetal adducts which are formed in spontaneous reactions between glutathionyl groups and aldehydes. The primary physiological function of glyoxalase I is the detoxification of methylglyoxal, a reactive 2-oxoaldehyde that is cytostatic at low concentrations and cytotoxic at millimolar concentrations. Glyoxalase I is a target for the development of pharmaceuticals against bacteria, protozoans and human cancer.\n\nSimple, direct and automation-ready procedures for measuring GLO-1 activity in biological samples are highly desirable in research and drug discovery. Glyoxalase I assay kit provides a sensitive and convenient method for GLO-1 activity determination. The method involves monitoring the increase in the product of the GLO-1 reaction, Slactoylglutathione, by measuring the change in absorbance at 240 nm.\n\nKey Features:\nSensitive and accurate. Detection limit 4 U/L GLO-1 activity.\nSimple and high-throughput. The procedure involves incubation of the provided substrate with the sample in a microplate. Can be readily automated as a high-throughput assay for thousands of samples per day.\n\nApplications:\nDirect Assays: GLO-1 activity in enzyme preparations or biological samples.\nDrug Discovery/Pharmacology: effects of drugs on GLO-1 activity.\n\nKit Contents:\nAssay Buffer (pH 6.6): 20ml Substrate: 1ml\n96 well UV Titer Plate 1 Plate Cosubstrate: 1ml\nStorage conditions. Kit is shipped at room temperature. Store the plate at room temperature and other components at -20 degrees C. Shelf life: 12 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\nProcedure For Pure Enzyme Preparations:\n1. Bring all reagents to room temperature prior to assay.\n2. Add 40ul of each sample to separate wells of the 96 well UV titer plate. Also, include one blank (sample buffer without GLO-1) well per assay run.\n3. Prepare Working Reagent for all wells by mixing per well: 160ul Assay Buffer with 8ul Substrate and 8ul Cosubstrate. Add 160ul Working Reagent to each well.\n4. Read the optical density at 240 nm at t=0 min and again at t=10 min. If measuring low GLO-1 activities, longer reaction times can be used.\n\nProcedure For Proteinous Samples:\n1. Each sample requires 2 tubes: one for the GLO-1 Reaction and one for the Sample Blank. Add 40ul of each sample (serum samples should be diluted at least 2

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SPECIFICATIONS

Catalog Number

G8235-30E

Size

1Kit

References

1. Davis, KA and Williams, GR. (1969) Glyoxalase I, a lyase or an oxidoreductive isomerase? Can. J. Biochem 47: 553-6. 2. Ditzen, C et al. (2006) Protein biomarkers in a mouse model of extremes in trait anxiety. Mol Cell Proteomics 5: 1914-20. 3. Strzinek, RA. et al. (1972) The purification and characterization of liver glyoxalase I from normal mice and from mice bearing a lymphosarcoma. Cancer Res 32:2359-64.