Phosphate Assay Kit (Colorimetric) (ab65622)
Key features and details
- Assay type: Enzyme activity
- Detection method: Colorimetric
- Platform: Microplate reader
- Assay time: 2 hr
- Sample type: Cell culture media, Cell Lysate, Other biological fluids, Plasma, Serum, Tissue Extracts
- Sensitivity: 1 µM
Overview
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Product name
Phosphate Assay Kit (Colorimetric)
See all Phosphate kits -
Detection method
Colorimetric -
Sample type
Serum, Plasma, Other biological fluids, Tissue Extracts, Cell Lysate, Cell culture media -
Assay type
Enzyme activity -
Sensitivity
> 1 µM -
Range
0.001 mM - 1 mM -
Assay time
2h 00m -
Product overview
Phosphate Assay Kit (Colorimetric) (ab65622) provides an easy, quick and simple method for measuring phosphate levels.
The phosphate assay protocol uses a proprietary formulation of malachite green and ammonium molybdate which forms a chromogenic complex with phosphate ion giving an intense absorption band around OD = 650nm.
Phosphate assay protocol summary:
- add reaction mix to sample and standard wells
- incubate for 30 min
- analyze with a microplate reader -
Notes
This assay can be used with biological fluids but also inorganic samples such as algal blooms and water from run-off areas of high fertilizer use.
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Platform
Microplate reader
Properties
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Storage instructions
Store at room temperature. Please refer to protocols. -
Components 500 tests Phosphate Reagent 1 x 15ml Phosphate Standard (10 mM) 1 x 500µl -
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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Immunoprecipitates of Flag-NT5C ectopically expressed in HEK293 cells were incubated with 5 mM of the indicated nucleotides. Phosphate release was measured using a malachite green colorimetric assay (ab65622) and expressed as a percent of total nucleotide. The experiment was performed in duplicate and repeated 3 times independently. Error bars are sem.
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Standard curve: mean of duplicates (+/- SD) with background reads subtracted
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Phosphate measured in tissue lysates showing quantity (µmol) per mg of extracted protein.
Protein concentration for samples varied from 9 mg/mL to 14 mg/mL. Samples were diluted 400-4000 fold.
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Phosphate measured in cell culture lysates showing quantity (µmol) per mg of extracted protein.
Samples with the concentration of 1.3e7 cells/mL were used. Samples were diluted 400-4000 fold.
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Phosphate measured in biological fluids showing quantity (µmol) per mL of tested sample. Samples were diluted 100-1000 fold.