LS-K307-250
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bSLSBio LifeSpan BioSciences, Inc. Alkaline Phosphatase (AP) Assay Kit (Colorimetric) LS-K307-250 (250 Tests) • See Storage Conditions Below Introduction Alkaline phosphatase (ALP) catalyzes the hydrolysis of phosphate esters in an alkaline environment, resulting in the formation of an organic radical and inorganic phosphate. In mammals, this enzyme is found mainly in the liver and bones. Marked increase in serum ALP levels, a disease known as hyperalkalinephosphatasemia, has been associated with malignant biliary obstruction, primary biliary cirrhosis, primary sclerosing cholangitis, hepatic lymphoma and sarcoidosis. Simple, direct and automation-ready procedures for measuring ALP activity in serum are becoming popular in Research and Drug Discovery. This Alkaline Phosphatase Assay Kit is designed to measure ALP activity directly in biological samples without pretreatment. The improved Method utilizes p-nitrophenyl phosphate that is hydrolyzed by ALP into a yellow colored product (maximal absorbance at 405nm). The rate of the reaction is directly proportional to the enzyme activity. ALP p-Nitiophenyl phosphate-p-nitophenol + phosphate Key Features • High sensitivity and wide linear range. Use 5 pL serum or plasma sample. The detection limit is 2 U/L, linear up to 800 U/L. • Homogeneous and simple procedure. Simple "mix-and-measure" procedure allows reliable quantitation of ALP activity within 5 minutes. • Robust and amenable to HTS. All reagents are compatible with high-throughput liquid handling instruments. Applications • Direct Assays: ALP activity in serum, plasma and other sources. • Characterization and Quality Control for ALP production. • Drug Discovery: high -throughput screen for ALP inhibitors and evaluation of ALP inhibitors. Materials Not Supplied Pipetting devices and accessories (e.g. multi-channel pipettor). Procedure using 96-well plate: Clear bottom 96-well plates (e.g. Corning Costar) and plate reader. Procedure using cuvette: Spectrophotometer and cuvettes for measuring OD 405nm. FOR RESEARCH USE ONLY! Not for use in humans. LifeSpan BioSciences, Inc. • 2401 Fourth Avenue, Suite 900, Seattle, WA 98121 www.LSBio.com • (206) 464-1554 • TechnicalSupport@LSBio.com

bSLSBio LifeSpan BioSciences, Inc. Alkaline Phosphatase (AP) Assay Kit (Colorimetric) LS-K307-250 (250 Tests) • See Storage Conditions Below Storage The kit is shipped at room temperature. Store pNPP Liquid at -20°C and all other components at 4°C. Shelf life of 12 months after receipt. Assay Procedure This assay is based on a kinetic reaction. Use of a multi-channel pipettor is recommended. Addition of Working Reagent to samples should be quick and mixing should be brief but thorough. Assays can be executed at room temperature or 37°C. Reagent preparation: equilibrate reagents to room...

bSLSBio LifeSpan BioSciences, Inc. Alkaline Phosphatase (AP) Assay Kit (Colorimetric) LS-K307-250 (250 Tests) • See Storage Conditions Below Calculation ALP activity of the sample (IU/L) is _(ODsamplet- ODsamplec) - 1000 - Reaction Vol ~ f-e-I - Sample Vol = 266.7 X (ODsahple<-ODsajwleb) Note: (1) if sample ALP activity exceeds 800 IU/L, dilute samples in saline and repeat the assay, multiply the result by the dilution factor. (2) incubation can be prolonged for samples with low ALP activity. Sample Data Samples were assayed in duplicate (n = 2) using the 96-well plate protocol. The ALP...