Product Name: Acid Phosphatase Activity Colorimetric Assay 

Cat. #: T2075-100 

Size: 100 assays 

Ship at 4℃, 

Store at -20℃, 

Avoid light 

Shelf Life: 12monthsDescription

Acid phosphatases (AP) dephosphorylate phosphate groups from phosphate esters under acidconditions. Different acid phosphatase isozymes are found in different organs, and their serumlevels are usedasadiagnostic index for disease in the corresponding organs. Elevated prostatic acid phosphatase levels mayindicate the presence of prostate cancer and elevated tartrate-resistant acid phosphatase levels mayindicatebone disease. This Kit provides a high-sensitive, simple, and direct assay approach to measure APactivityinserum and other samples. It is suitable for research and drug discovery. The kit uses p-nitrophenyl phosphate(pNPP) as a phosphatase substrate which turns yellow (λmax = 405 nm) when dephosphorylatedbyAP. Thekitcan detect as low as 20 µU acid phosphatase activity in samples. Applications Direct Assays: Acid phosphatase in serum, plasma, urine, and other bio-samples. Key Features

Flexible: Suitable for colorimetric assay. Accurate: Use 50 µL samples. Detection ranges from 0.4-200 µU in a 96-well plate for colorimetric assay. Simple and high-throughput: Just load-incubate-Read. The kit can be used for a robust method. Fast: less than 30 minutes. Kit Component

Assay Buffer: 10mL Substrate: 0.5mL Enzyme Standard Stock (2U/mL): 100 µL Stop Solution: 10mLPrecaution

Inhibitors of AP, such as tartrate, fluoride, EDTA, oxalate, and citrate, should be avoided in samplepreparation.Sample Preparations

Sample Preparations: Serum, plasma, urine, semen, and cell culture media can be assayed directly. Cells (1×105 ) or tissue (~10 mg) can be homogenized in 150 µl Assay Buffer, centrifuge to removeinsolublematerial at 13,000g, 3 minutes. The supernatant can be used as test sample for the assay testing. Standard Curve Preparations (Fig. 1)

1. Label 1.5mL tube from Std1 to 8. As below the diagram.

 2. Add 360 µL of 1x Assay Buffer to Std1, and 200 µL to Std2 to 8. 

3. Take 40 µL of 1U/mL AP Standard Stock solution to Std1, then make 2x series dilution fromStd2throughStd7 by transferring 200µL to the next concentration, Std8 is 1x Assay Buffer alone as a standard0. 

The standard concentration range is 200, 100, 50, 25, 12.5, 6.25,3.125 µM, and 0.

Assay Procedures

1. Add 45 µL of standard or sample to each well of a microplate in duplicate manner. 2. Add 5 µL substrate to each well, and incubate at room temperature for 15-60 minutes, protect fromlight.3. Add50 µL Stop Solution to terminate the reaction, and fully mix. 4. Measure OD value at test wavelength of 405 nm, and a reference wavelength of 630 nmin a plater reader.Related Products

ATP Colorimetric/Fluorometric Assay (T2010) Glucose Oxidase Colorimetric/Fluorometric Assay(T2088)ADP Colorimetric/Fluorometric Assay Kit (T2020) β-Hexosaminidase Activity Assay (T2105)

Cytochrome C Oxidase Activity Assay (T2115)

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