Phosphate Assay Kit (Colorimetric - UV absorption) (ab219938) has been developed for measuring the activity of any phosphate-generating enzyme such as ATPases or phosphatases.
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- Journal of colloid and interface science 629:541-5532022Nanoarchitectonics of Layer-by-Layer (LbL) coated nanostructured lipid carriers (NLCs) for Enzyme-Triggered charge reversal.Applications:Unspecified applicationReactive species:Unspecified reactive speciesZeynep Burcu Akkuş-Dağdeviren,Andrea Fürst,Julian David Friedl,Martina Tribus,Andreas Bernkop-SchnürchPubMed 36088699
Key facts
- Detection method
- Colorimetric
- Sample types
- Tissue Lysate, Urine, Plasma, Serum, Other biological fluids, Cell Lysate
- Assay type
- Quantitative
- Reactive species
- Mammals
- Sensitivity
- = 0.2 µM
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Phosphate Assay Kit (Colorimetric - UV absorption) (ab219938) has been developed for measuring the activity of any phosphate-generating enzyme such as ATPases or phosphatases.
Key facts
- Detection method
- Colorimetric
- Sample types
- Tissue Lysate, Urine, Plasma, Serum, Other biological fluids, Cell Lysate
- Assay type
- Quantitative
- Reactive species
- Mammals
- Assay Platform
- Microplate reader
- Sensitivity
- = 0.2 µM
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- -20°C
- Appropriate long-term storage conditions
- -20°C
- Storage information
- -20°C
Notes
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.
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Phosphate a simple inorganic chemical functions mechanically as a component of various essential biological molecules such as nucleic acids ATP and phospholipids. Phosphate ions serve as an important player in cellular energy transfer through involvement in phosphorylation reactions. It's present abundantly in living organisms especially in the form of calcium phosphate in bones and teeth. The molecular weight of a phosphate ion is approximately 94.97 g/mol. Expressed ubiquitously across different tissues and cells phosphate is also available in multiple assay formats like the phosphate assay kit and phosphorus assay kits for scientific research.
- Biological function summary
Phosphate plays a role in numerous cellular processes beyond its participation in energy metabolism. It contributes critically to cellular signaling pathways as a part of second messengers like cyclic AMP and is a core part of protein kinase and phosphatase complexes which regulate many cellular functions. Moreover phosphate acts in buffering systems maintaining cellular pH balance. In the cellular context the probe and colorimetric malachite green assay often quantify phosphate levels aiding in understanding phosphate homeostasis in biochemical studies.
- Pathways
The chemical phosphate is integral to significant pathways such as glycolysis and the citric acid cycle where it assists in energy production and storage. It associates with proteins like hexokinase in the glycolytic pathway and is critical for the formation of high-energy the phosphate bonds in ATP. As phosphate cycles through these pathways it continuously modulates various metabolic activities and biochemical reactions within the cell highlighting its universal importance.
- Associated diseases and disorders
Phosphate imbalances relate to conditions such as chronic kidney disease and hypophosphatemia. In chronic kidney disease altered phosphate homeostasis can lead to cardiovascular complications due to phosphate's interactions with proteins such as fibroblast growth factor 23 (FGF23) which regulates phosphate metabolism. In hypophosphatemia a deficiency in phosphate can result in skeletal disorders highlighting the need for precise phosphate detection and monitoring in clinical settings.
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Functional Studies - 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.
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