Malachite Green Phosphate Detection Kit

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Product Details
Citations (25)
Reviews (2)

Malachite Green Phosphate Detection Kit Summary

Assay Type
Malachite Green Phosphate Detection Kit
Assay Length
4 hours 40 minutes (after plate preparation)
Sample Volume Required
50 µL
Sufficient Materials
For fifteen 96-well plates*
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

Product Features

This Malachite Green Phosphate Detection Kit employs a simple, sensitive, reproducible, and non-radioactive method for measuring inorganic phosphate in aqueous solutions. The assay is based on the malachite green-molybdate binding reaction, and has a wide variety of applications, including the measurement of protein and lipid phosphorylation and phosphatase activity. The protocol presented is for the assay of phosphate in a 96-well microplate format. The Malachite Green Phosphate Detection Kit can also be run using 1 mL cuvettes or 384-well plates suitable for high-throughput screening.

Kit Content

  • Malachite Green Reagent A - Ammonium molybdate in 3M sulfuric acid
  • Malachite Green Reagent B - Malachite green oxalate and polyvinyl alcohol
  • Phosphate Standard

Other Reagents Required

Microplates: R&D Systems (Catalog # DY990), or equivalent

Plate Sealers: ELISA Plate Sealers (Catalog # DY992), or equivalent


Product Datasheets

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Preparation and Storage

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at room temperature. Do not use past expiration date.

Citations for Malachite Green Phosphate Detection Kit

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

25 Citations: Showing 1 - 10
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  1. Phosphate uptake restriction, phosphate export, and polyphosphate synthesis contribute synergistically to cellular proliferation and survival
    Authors: Takado, M;Komamura, T;Nishimura, T;Ohkubo, I;Ohuchi, K;Matsumoto, T;Takeda, K;
    The Journal of biological chemistry
    Species: S. pombe
    Sample Types: Cell Extract, Cell Lysates
  2. Adjustment of light-responsive NADP dynamics in chloroplasts by stromal pH
    Authors: Fukuda, Y;Ishiyama, C;Kawai-Yamada, M;Hashida, SN;
    Nature communications
    Species: A. thaliana
    Sample Types: Cell Lysates
  3. Human soluble CD39 displays substrate inhibition in a substrate-specific manner
    Authors: Vadlamani, VMK;Gunasinghe, KKJ;Chee, XW;Rahman, T;Harper, MT;
    Scientific reports
    Species: Human
    Sample Types: Recombinant Protein
  4. Anopheles salivary apyrase regulates blood meal hemostasis and drives malaria parasite transmission
    Authors: Pala, ZR;Alves E Silva, TL;Minai, M;Crews, B;Patino-Martinez, E;Carmona-Rivera, C;Valenzuela-Leon, PC;Martin-Martin, I;Flores-Garcia, Y;Cachau, RE;Srivastava, N;Moore, IN;Alves, DA;Kaplan, MJ;Fischer, E;Calvo, E;Vega-Rodriguez, J;
    bioRxiv : the preprint server for biology
  5. PI3K drives the de novo synthesis of coenzyme A from vitamin B5
    Authors: CC Dibble, SA Barritt, GE Perry, EC Lien, RC Geck, SE DuBois-Coy, D Bartee, TT Zengeya, EB Cohen, M Yuan, BD Hopkins, JL Meier, JG Clohessy, JM Asara, LC Cantley, A Toker
    Nature, 2022-07-27;608(7921):192-198.
    Species: Human
    Sample Types: Proteins
  6. Mechanistic Roles of Tyrosine Phosphorylation in Reversible Amyloids, Autoinhibition, and Endosomal Membrane Association of ALIX
    Authors: RD Elias, B Ramaraju, L Deshmukh
    The Journal of Biological Chemistry, 2021-10-22;0(0):101328.
    Species: Human
    Sample Types: Recombinant Protein
  7. A Novel CD73 Inhibitor SHR170008 Suppresses Adenosine in Tumor and Enhances Anti-Tumor Activity with PD-1 Blockade in a Mouse Model of Breast Cancer
    Authors: S Liu, D Li, J Liu, H Wang, I Horecny, R Shen, R Zhang, H Wu, Q Hu, P Zhao, F Zhang, Y Yan, J Feng, L Zhuang, J Li, L Zhang, W Tao
    OncoTargets and Therapy, 2021-08-24;14(0):4561-4574.
    Species: Human, Mouse
    Sample Types: Protein, Recombinant Proteins, Whole Cells
  8. The acid-resistance potential of arginine-fluoride varnish treated enamel
    Authors: MN Bijle, MM Abdalla, U Ashraf, KW Siu, J Tsoi, CKY Yiu
    Journal of the Mechanical Behavior of Biomedical Materials, 2021-08-08;125(0):104763.
    Species: Human
    Sample Types: Complex Sample Type
  9. Regulation of inorganic polyphosphate is required for proper vacuolar proteolysis in fission yeast
    Authors: N Sawada, S Ueno, K Takeda
    The Journal of Biological Chemistry, 2021-06-18;0(0):100891.
    Species: Fission Yeast
    Sample Types: Cell Lysates
  10. The effect of arginine-fluoride varnish on biochemical composition of multi-species biofilm
    Authors: MN Bijle, U Ashraf, MM Abdalla, CKY Yiu
    Journal of dentistry, 2021-03-09;108(0):103631.
    Species: Human
    Sample Types: Biofilm
  11. The enamel remineralization potential of fluoride varnishes containing arginine
    Authors: MN Bijle, M Ekambaram, EC Lo, CKY Yiu
    J Dent, 2020-06-22;0(0):103411.
    Species: Human
    Sample Types: Complex Sample Type
  12. The C-terminal domain of feline and bovine SAMHD1 proteins has a crucial role in lentiviral restriction
    Authors: C Wang, K Zhang, L Meng, X Zhang, Y Song, Y Zhang, Y Gai, Y Zhang, B Yu, J Wu, S Wang, X Yu
    J. Biol. Chem., 2020-02-19;0(0):.
    Species: Human
    Sample Types: Whole Cells
  13. Unfolding of a ClC chloride transporter retains memory of its evolutionary history
    Authors: D Min, RE Jefferson, Y Qi, JY Wang, MA Arbing, W Im, JU Bowie
    Nat. Chem. Biol., 2018-03-26;0(0):.
  14. Green Tea Polyphenol EGCG Upregulates Tollip Expression by Suppressing Elf-1 Expression
    Authors: M Kumazoe, M Yamashita, Y Nakamura, K Takamatsu, J Bae, S Yamashita, S Yamada, H Onda, T Nojiri, K Kangawa, H Tachibana
    J. Immunol., 2017-09-27;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  15. CD73 promotes resistance to HER2/ErbB2 antibody therapy
    Authors: M Turcotte, D Allard, D Mittal, Y Bareche, L Buisseret, V Jose, S Pommey, V Delisle, S Loi, H Joensuu, PL Kellokumpu, C Sotiriou, MJ Smyth, J Stagg
    Cancer Res., 2017-08-30;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  16. ADPase CD39 Fused to Glycoprotein VI-Fc Boosts Local Antithrombotic Effects at Vascular Lesions
    Authors: H Degen, O Borst, M Ziegler, AK Mojica Mun, J Jamasbi, B Walker, S Göbel, J Fassbender, K Adler, R Brandl, G Münch, R Lorenz, W Siess, M Gawaz, M Ungerer
    J Am Heart Assoc, 2017-07-27;6(8):.
    Species: Mouse
    Sample Types: Plasma
  17. Human alkaline phosphatase dephosphorylates microbial products and is elevated in preterm neonates with a history of late-onset sepsis
    Authors: M Pettengill, JD Matute, M Tresenrite, J Hibbert, D Burgner, P Richmond, J Luis Millá, A Ozonoff, T Strunk, A Currie, O Levy
    PLoS ONE, 2017-04-27;12(4):e0175936.
    Species: Human
    Sample Types: Recombinant Protein
  18. Tiamulin inhibits breast cancer growth and pulmonary metastasis by decreasing the activity of CD73
    Authors: X Yang, S Pei, H Wang, Y Jin, F Yu, B Zhou, H Zhang, D Zhang, D Lin
    BMC Cancer, 2017-04-11;17(1):255.
    Species: Human, Mouse
    Sample Types: Whole Cells
  19. Bone marrow-derived mesenchymal stromal cells harness purinergenic signaling to tolerize human Th1 cells in vivo.
    Authors: Amarnath S, Foley J, Farthing D, Gress R, Laurence A, Eckhaus M, Metais J, Rose J, Hakim F, Felizardo T, Cheng A, Robey P, Stroncek D, Sabatino M, Battiwalla M, Ito S, Fowler D, Barrett A
    Stem Cells, 2015-04-01;33(4):1200-12.
    Species: Human
    Sample Types: Whole Cells
  20. Enzymatic basis for N-glycan sialylation: structure of rat alpha2,6-sialyltransferase (ST6GAL1) reveals conserved and unique features for glycan sialylation.
    Authors: Meng L, Forouhar F, Thieker D, Gao Z, Ramiah A, Moniz H, Xiang Y, Seetharaman J, Milaninia S, Su M, Bridger R, Veillon L, Azadi P, Kornhaber G, Wells L, Montelione G, Woods R, Tong L, Moremen K
    J Biol Chem, 2013-10-23;288(48):34680-98.
    Species: Rat
    Sample Types: Protein, Whole Cells
  21. CD39/adenosine pathway is involved in AIDS progression.
    Authors: Nikolova M, Carriere M, Jenabian MA, Limou S, Younas M, Kok A, Hue S, Seddiki N, Hulin A, Delaneau O, Schuitemaker H, Herbeck JT, Mullins JI, Muhtarova M, Bensussan A, Zagury JF, Lelievre JD, Levy Y
    PLoS Pathog., 2011-07-07;7(7):e1002110.
    Species: Human
    Sample Types: Cell Lysates
  22. NCAM induces CaMKIIalpha-mediated RPTPalpha phosphorylation to enhance its catalytic activity and neurite outgrowth.
    Authors: Bodrikov V, Sytnyk V, Leshchyns'ka I, den Hertog J, Schachner M
    J. Cell Biol., 2008-09-22;182(6):1185-200.
  23. Protein cysteine sulfinic acid reductase (sulfiredoxin) as a regulator of cell proliferation and drug response.
    Authors: Lei K, Townsend DM, Tew KD
    Oncogene, 2008-05-05;27(36):4877-87.
    Species: Human
    Sample Types: Cell Lysates
  24. Moderate consumption of red wine (cabernet sauvignon) improves ischemia-induced neovascularization in ApoE-deficient mice: effect on endothelial progenitor cells and nitric oxide.
    Authors: Lefevre J, Michaud SE, Haddad P, Dussault S, Menard C, Groleau J, Turgeon J, Rivard A
    FASEB J., 2007-07-19;21(14):3845-52.
    Species: Human
    Sample Types: Cell Lysates
  25. Akt-mediated activation of HIF-1 in pulmonary vascular endothelial cells by S-nitrosoglutathione.
    Authors: Carver DJ, Gaston B, Deronde K, Palmer LA
    Am. J. Respir. Cell Mol. Biol., 2007-05-31;37(3):255-63.
    Species: Bovine
    Sample Types: Cell Lysates


  1. Does the Malachite Green Phosphate Detection Kit (DY996) detect both pyrophosphate (diphosphate) and free phosphate (monophosphate)?

    • The Malachite Green Phosphate Detection Kit measures free inorganic phosphate in aqueous solutions. The assay principle is based on complex formation between malachite green molybdate and free phosphate under acidic conditions. Lipid bound, protein-bound, pyrophashate and other condensed phosphates must be hydrolyzed prior to measurement in the malachite green assay.

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Reviews for Malachite Green Phosphate Detection Kit

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Malachite Green Phosphate Detection Kit
By Anonymous on 09/21/2023
Sample Tested: Recombinant protein

Attached is a picture of the standard curve in assay buffer, and the Raw OD readings. Some small variation but still a solid R^2. The Assay buffer includes 3mM DTT and 100mM ATP with no adverse effects observed. Nice alternative to radioactive P32 assays and temperature-sensitive LDH kits.

Malachite Green Phosphate Detection Kit
By Anonymous on 09/07/2017
Sample Tested: Recombinant protein