Phosphate Assay Kit (Colorimetric - UV absorption) (ab219938)
Key features and details
- Assay type: Quantitative
- Detection method: Colorimetric
- Platform: Microplate reader
- Sample type: Cell Lysate, Other biological fluids, Plasma, Serum, Tissue Lysate, Urine
- Sensitivity: 0.2 µM
Overview
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Product name
Phosphate Assay Kit (Colorimetric - UV absorption)
See all Phosphate kits -
Detection method
Colorimetric -
Sample type
Urine, Serum, Plasma, Other biological fluids, Cell Lysate, Tissue Lysate -
Assay type
Quantitative -
Sensitivity
0.2 µM -
Species reactivity
Reacts with: Mammals, Other species -
Product overview
Phosphate Assay Kit (Colorimetric - UV absorption) (ab219938) has been developed for measuring the activity of any phosphate-generating enzyme such as ATPases or phosphatases. In the presence of inorganic phosphate, the MESG reagent is converted to 2-amino-6-mercapto-7-methlpurine by purine nucleoside phosphorylase with absorption wavelength shift to red. This feature has been used to develop our convenient MESG-based phosphate assay kit, an alternative to hazardous radioactive methods. The MESG substrate gives an absorbance increase at OD 360 nm on phosphorylysis at pH 6.5-8.5. The assay is shown to quantitate phosphate at the final concentration as low as 0.2 µM in cell lysates or biological fluids.
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Notes
Phosphate is involved in many biological processes. For example, phosphatases, ATPases and several other enzymes catalyze biochemical reactions in which inorganic phosphate (Pi) is released from a phosphoester substrate. The detection of many phosphoester–metabolizing enzymes is difficult because suitable substrates are not available. It is usually necessary to determine inorganic phosphate release using tedious colorimetric assays or radioisotope based methods.
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Platform
Microplate reader
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 200 tests 1 mM KH2PO4 1 x 1ml Assay Buffer 1 x 10ml MESG Substrate (10 mg) 1 vial Purine Nucleoside Phosphorylase (1 mg) 1 vial -
Research areas
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Relevance
Phosphate is one of the most important of the inorganic ions in biological systems. It functions in a variety of roles. One of the most important roles is as a molecular switch, turning enzyme activity on and off through the mediation of the various protein kinases and phosphatases in biological systems. Phosphate is also of great importance in mineralization processes and is a primary stimulus of algal blooms frequently found in bodies of fresh water, due to run-off from areas of high fertilizer use.
Images
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Phosphate Assay Kit (Colorimetric - UV absorption) (ab219938)
Typical Phosphate standard calibration curve. Phosphate dose response was measured on a 96 well UV plate using a SpectraMax Plus microplate reader (Molecular Devices). As low as 0.2 µM phosphate can be detected with 30 minutes incubation.
