PhosphoWorks™ Colorimetric Phosphate Assay Kit  *Blue Color*

Phosphate is involved in many biological reactions. For example, phosphatases, ATPases and several other enzymes catalyze reactions in which inorganic phosphate (Pi) is released from a substrate. This PhosphoWorks™ Phosphate Assay Kit has been developed for measuring the activity of any Pi-generating enzyme. The kit is formulated to give sensitive detection of Pi, providing an alternative to hazardous radioactive methods and other less sensitive colorimetric assays. The measurement of Pi is based on the change in absorbance of a malachite green derivative in the presence of molybdate. Unlike other malachite dye formulations, this kit gives a completely stable end-point signal that is not prone to precipitation.


Kit Features

Broad Applications : Can be used for monitoring any biological processes that either generate or consume phosphate.
Convenient : Formulated to have minimal hands-on time.
Non-Radioactive : No special requirements for waste treatment.
Use of Native substrates : Substrates can be proteins, peptides, nucleotides, sugars, organic molecules or inorganic
                                     salts.

Kit Components

Component A : 1 mM KH2PO4 1 vial (1 mL)                  4 °C
Component B : MG PlusTM Reagent 1 bottle (20 mL)    4 °C and avoid light

Brief Summary

→ Prepare test samples (80 μL) along with phosphate standard dilutions (80 μL) from component A
→ Add component B (20 μL)
→ Incubate at room temperature for 10-40 min
→ Read Absorbance at 600-660 nm

References

1. Bernal C, Palacin C, Boronat A, Imperial S. (2005) A colorimetric assay for the determination of 4-diphosphocytidyl-2-C-methyl-D-erythritol 4-phosphate synthase activity. Anal Biochem, 337, 55.
2. Hannig C, Hamkens A, Becker K, Attin R, Attin T. (2005) Erosive effects of different acids on bovine enamel: release of calcium and phosphate in vitro. Arch Oral Biol, 50, 541.
3. Mahuren JD, Coburn SP, Slominski A, Wortsman J. (2001) Microassay of phosphate provides a general method for measuring the activity of phosphatases using physiological, nonchromogenic substrates such as lysophosphatidic acid. Anal Biochem, 298, 241.
4. Cala SE. (1999) Determination of a putative phosphate-containing peptide in calreticulin. Biochem Biophys Res Commun, 259, 233.