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Phosphate Assay Kit, BioAssay(TM), Colorimetric

Cat no: P4071-72T

Phosphate Assay Kit, BioAssay(TM), Colorimetric

The Phosphate Assay Kit is based on a proprietary formulation of the malachite green dye. The reagent forms a blue colored complex with free orthophosphate. The rapid color formation from the reaction can be conveniently measured on a spectrophotometer (600-660 nm) or on a plate reader. The non-radioactive Colorimetric Assay: kits have been optimized to offer superior sensitivity and prolonged shelf life. The assay is simple and fast, involving a single addition step for phosphate determination. Assays can be performed in tubes, cuvettes or multi-well plates. The assays can be conveniently executed in 96-well plates for high-throughput screening of enzyme inhibitors.\n\nKey Features:\nReagent very stable. Due to our innovative formulation, no precipitation of reagent occurs. Therefore no filtration of reagent is needed prior to assays, as is often required with other commercial kits.\nHigh sensitivity and wide detection range: detection of as little of 20pmoles of phosphate and useful range between 0.4uM and 50uM phosphate.\nFast and convenient: single reagent

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SPECIFICATIONS

Catalog Number

P4071-72T

Size

1Kit

References

1. Rumsfeld J, Ziegelbauer K, Spaltmann F (2000). High-throughput assay for inorganic pyrophosphatases using the cytosolic enzymes of Saccharomyces cerevisiae and human as an example. Protein Expr Purif. 18(3): 303-9.\n2. Cogan EB, Birrell GB, Griffith OH (1999). A robotics-based automated assay for inorganic and organic phosphates. Anal Biochem. 271: 29-35.\n3. Ng DH, Harder KW, Clark-Lewis I, Jirik F, Johnson P (1995). Non- radioactive method to measure CD45 protein tyrosine phosphatase activity isolated directly from cells. J Immunol Methods. 179(2): 177-85.\n4. Fisher DK, Higgins TJ (1994). A sensitive, high-volume, colorimetric assay for protein phosphatases. Pharm Res. 11(5): 759-63.\n5. Harder KW, Owen P, Wong LK, Aebersold R, Clark-Lewis I, Jirik FR (1994). Characterization and kinetic analysis of the intracellular domain of human protein tyrosine phosphatase beta (HPTP beta) using synthetic phosphopeptides. Biochem J. 298(2): 395-401.\n6. Queiroz-Claret C, Meunier JC (1993). Staining technique for phosphatases in polyacrylamide gels. Anal Biochem. 209(2): 228-31.\n7. Geladopoulos TP, Sotiroudis TG, Evangelopoulos AE (1991). A malachite green colorimetric assay for protein phosphatase activity. Anal Biochem. 192(1): 112-6.\n8. Samizo K, Ishikawa R, Nakamura A, Kohama K (2001). A highly sensitive method for measurement of myosin ATPase activity by reversed- phase high-performance liquid chromatography. Anal Biochem. 293: 212-5.\n9. Hackney DD, Jiang W (2001). Assays for kinesin microtubule-stimulated ATPase activity. Methods Mol Biol. 164: 65-71.\n10. Cen X, Huang Y, Wang R, Chen Z, Wu Z (1998). Simultaneous assay of Ca(2+)-ATPase and Na+, K(+)-ATPase activities of osteoblast rat by malachite green colorimetic method. Hua Xi Yi Ke Da Xue Xue Bao. 29(4): 427-30.\n11.Mahuren JD, Coburn SP, Slominski A, Wortsman J (2001). Microassay of phosphate provides a general method for measuring the activity of phosphatases using physiological, nonchromogenic substrates such as lysophosphatidic acid. Anal Biochem. 298(2): 241-5.\n12. Cala SE (1999). Determination of a putative phosphate-containing peptide in calreticulin. Biochem Biophys Res Commun. 259(2): 233-8.\n13. Gibson NJ, Newton CR, Little S (1997). A colorimetric assay for phosphate to measure amplicon accumulation in polymerase chain reaction. Anal Biochem. 254(1): 18-22.\n14. Ekman P, Jager O (1993). Quantification of subnanomolar amounts of phosphate bound to seryl and threonyl residues in phosphoproteins using alkaline hydrolysis and malachite green. Anal Biochem. 214: 138-41.\n15. Lu X, Pearson A, Lunec J (2003). The MYCN oncoprotein as a drug development target. Cancer Lett. 197(1-2): 125-30.\nClinical and General Diagnosis\n16. Ohyama T, Matsubara C, Takamura K (1996). Sensitive densitometry for the determination of platelet-activating factor and other phospholipids in human tears. Analyst. 121(12): 1943-7.\n17. Matsubara C, Ohyama T, Takamura K (1994). Densitometric quantitation of platelet activating factor and other phospholipids in human saliva using enzyme reaction on a silica plate. Yakugaku Zasshi. 114(9): 681-90.\n18. Kirchgesser M, Dahlmann N (1990). A colorimetric assay for the determination of acid nucleoside triphosphatase activity. J Clin Chem Clin Biochem. 28(6): 407-11.\n19. D'Angelo E, Crutchfield J, Vandiviere M (2001). Rapid, sensitive, microscale determination of phosphate in water and soil. J Environ Qual. 30(6): 2206-2209.

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