Malachite Green Phosphate Assay Kit
Protocol SDS  Printed Protocols and SDSs are not provided with kits.

For sensitive and high-throughput phosphate determination.

Key Features
Reagent very stable. Due to our innovative formulation, no precipitation of reagent occurs. Therefore no filtration of reagent is needed prior to assays, as is often required with other commercial kits.

High sensitivity and wide detection range: detection of as little of 1.6 pmoles of phosphate and useful range between 0.02 μM and 40 μM phosphate.

Fast and convenient: homogeneous "mix-and-measure" assay allows quantitation of free phosphate within 20 minutes.

Compatible with routine laboratory and HTS formats: assays can be performed in tubes, cuvettes or microplates, on spectrophotometers and plate readers.

Robust and amenable to HTS: Z factors of 0.7 to 0.9 are observed in 96-well and 384-well plates. Can be readily automated on HTS liquid handling systems.

OD620nm (malachite green)

Biological, environment etc


30 min

2500 tests

Detection Limit
0.02 μM

Shelf Life
12 months

More Details
The Malachite Green Phosphate Assay Kit is based on quantification of the green complex formed between Malachite Green, molybdate and free orthophosphate. The rapid color formation from the reaction can be conveniently measured on a spectrophotometer (600 - 660 nm) or on a plate reader. The non-radioactive colorimetric assay kits have been optimized to offer superior sensitivity and prolonged shelf life. The assay is simple and fast, involving a single addition step for phosphate determination. Assays can be executed in tubes, cuvettes or multi-well plates. The assays can be conveniently performed in 96- and 384-well plates for high-throughput screening of enzyme inhibitors.

1. What is the difference between your 3 phosphate assays: DIPI-500, POMG-25H, POPB-500?

All three products are based on the malachite green dye. While the PiBlue (POPB-500) and QuantiChrom (DIPI-500) Phosphate Assay Reagents are identical, they differ in the standards. The PiBlue is designed for customers who want prepare a full calibration curve for their assays, and the latter has one set of a blank control and 30 µM phosphate standard. The reagent is more diluted than the reagent used in the Malachite Green Phosphate Assay Kit (POMG-25H). As a result, one adds 100 µL reagent (POPB-500 or DIPI-500) to 50 µL sample, whereas for the Malachite Green based assay, one adds 20 µL Reagent to 80 µL sample.

In practice, the DIPI-500 is the most convenient kit to use, because there is no need to prepare a calibration curve there is no need for extra mixing. Because adding a larger volume (100 μL) to a small volume (50 μL) effects efficient mixing in well, no extra mixing is needed. The POPB-500 requires preparation of calibration curve. With the POMG-25H Kit, one needs to mix the reagent with the sample, because adding 20 µL to 80 µL does not mix efficiently by itself. Also a preparation of a standard curve is necessary.

For more detailed product information and questions, please feel free to Contact Us. Or for more general information regarding our assays, please refer to our General Questions
1. Kim HE, et al (2005). Formation of apoptosome is initiated by cytochrome c-induced dATP hydrolysis and subsequent nucleotide exchange on Apaf-1. PNAS 102(49):17545-50. Assay: Phosphate in Human Enzyme (Pubmed).

2. Guérette, D. et al (2007). Molecular evolution of type VI intermediate filament proteins. BMC Evolutionary Biology 7:164. Assay: Human ATPase, GTPase (Pubmed).

3. Green, M.L. et al (2005). Ethylene glycol induces hyperoxaluria without metabolic acidosis in rats. Am J Physiol Renal Physiol 289: F536–F543. Assay: Phosphate in Rat Urine, Serum (Pubmed).

4. Saran, D. et al (2006). Multiple-turnover thio-ATP hydrolase and phospho-enzyme intermediate formation activities catalyzed by an RNA enzyme. Nucleic Acids Res. 34(11): 3201–3208. Assay: Phosphate in Human RNA (Pubmed).

5. Adkins, M.W. et al (2007). Chromatin Disassembly from the PHO5 Promoter Is Essential for the Recruitment of the General Transcription Machinery and Coactivators. Mol. Cell. Biol. 27: 6372–6382. Assay: Phosphate in Yeast Culture (Pubmed).

6. Guo W, et al (2006). The bioreduction of a series of benzoquinone ansamycins by NAD(P)H:quinone oxidoreductase 1 to more potent heat shock protein 90 inhibitors, the hydroquinone ansamycins. Mol Pharmacol. 70(4):1194-203. Assay: Phosphate in Yeast ATPase (Pubmed).

7. Lu B, et al (2007). Roles for the human ATP-dependent Lon protease in mitochondrial DNA maintenance. J Biol Chem. 282(24):17363-74. Assay: Phosphate in Human Enzyme (Pubmed).

8. Koroleva O, et al (2007). Structural conservation of RecF and Rad50: implications for DNA recognition and RecF function. EMBO J. 26(3):867-77. 17255941 Assay: Phosphate in Bacterial Enzyme (Pubmed).

9. Anand SP, et al (2007). DNA helicase activity of PcrA is not required for the displacement of RecA protein from DNA or inhibition of RecA-mediated strand exchange. J Bacteriol. 189(12):4502-9. Assay: Phosphate in Bacterial Enzyme (Pubmed).

10. Blumental-Perry A, et al (2006). Phosphatidylinositol 4-phosphate formation at ER exit sites regulates ER export. Dev Cell. 11(5):671-82. Assay: Phosphate in Rat Liposomes (Pubmed).

11. Guo W, et al (2005). Formation of 17-allylamino-demethoxygeldanamycin (17-AAG) hydroquinone by NAD(P)H:quinone oxidoreductase 1: role of 17-AAG hydroquinone in heat shock protein 90 inhibition. Cancer Res. 65(21):10006-15. Assay: Phosphate in Human Hsp90 (Pubmed).

12. Reigan P, et al (2011). A mechanistic and structural analysis of the inhibition of the 90-kDa heat shock protein by the benzoquinone and hydroquinone ansamycins. Mol Pharmacol. 79(5):823-32. Assay: Phosphate in Human Hsp90 (Pubmed).

13. Hsieh CW, et al (2010). Direct MinE-membrane interaction contributes to the proper localization of MinDE in E. coli. Mol Microbiol. 75(2):499-512. Assay: Bacterial ATPase, GTPase (Pubmed).

14. Liu et al (2011). Competition between foliar Neotyphodium lolii endophytes and mycorrhizal Glomus spp. fungi in Lolium perenne depends on resource supply and host carbohydrate content. Functional Ecology 25:910–920. Assay: Phosphate in Plant Extracts.

15. Yoshida C, et al (2010). Analysis of inhibition of topoisomerase IIalpha and cancer cell proliferation by ingenolEZ. Cancer Sci. 101(2):374-8. Assay: Phosphate in Human Topoisomerase (Pubmed).

16. Lin J, Wilson MA (2011). Escherichia coli thioredoxin-like protein YbbN contains an atypical tetratricopeptide repeat motif and is a negative regulator of GroEL. J. Biol Chem. 286(22):19459-69. Assay: Phosphate in Bacterial Enzyme (Pubmed).

17. Li Q, Uitto J (2010). The mineralization phenotype in Abcc6 ( -/- ) mice is affected by Ggcx gene deficiency and genetic background--a model for pseudoxanthoma elasticum. J Mol Med (Berl). 88(2):173-81. Assay: Phosphate in Mouse Tissue (Pubmed).

18. Cha C, et al (2011). Integrative design of a poly(ethylene glycol)-poly(propylene glycol)-alginate hydrogel to control three dimensional biomineralization. Biomaterials.32(11):2695-703. Assay: Phosphate in Hydrogels (Pubmed).

19. Aldana-Masangkay GI,et al (2011). Tubacin suppresses proliferation and induces apoptosis of acute lymphoblastic leukemia cells. Leuk Lymphoma. 52(8):1544-55. Assay: Phosphate in Human Cells (Pubmed).

20. Barisic S, et al (2008). Identification of PP2A as a crucial regulator of the NF-kappaB feedback loop: its inhibition by UVB turns NF-kappaB into a pro-apoptotic factor. Cell Death Differ. 15(11):1681-90. Assay: Phosphate in Human Cell Lysate (Pubmed).

21. Demidenko AA, et al (2011). Effects of viscogens on RNA transcription inside reovirus particles. J Biol Chem. 2011 Jun 30. Assay: Phosphate in Mouse Enzyme (Pubmed).

22. Soni, V et al (2015). Depletion of M. tuberculosis GlmU from Infected Murine Lungs Effects the Clearance of the Pathogen. PLoS Pathog 11(10):e1005235. Assay: Phosphate in Bacterial Enzyme (Pubmed).

To find more recent publications, please click here. To find more recent publications, please click here.
If you or your labs do not have the equipment or scientists necessary to run this assay, BioAssay Systems can perform the service for you.

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Please inquire via email or call 1-510-782-9988 x 2.
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Malachite Green Phosphate Assay Kit
Catalog No: POMG-25H
Price: $209    Qty:
For orders of 10 or more kits, please call 1-510-7829988x1 or email us for best pricing and/or bulk order.

Shipping: RT
Shipment: Fedex Service
Delivery: 1-2 days (US), 3-6 days (Intl) Storage: 4°C

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