PiBlue™ Phosphate Assay Kit

- Overview
  - The PiBlue™ Phosphate Assay Kit is based on a proprietary formulation of the malachite green dye. The PiBlue™ reagent forms a blue colored complex with 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 performed in tubes, cuvettes or multi-well plates. The assays can be conveniently executed in 96-well plates for high-throughput screening of enzyme inhibitors.
    
    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 20 pmoles of phosphate and useful range between 0.4 µM and 50 µM phosphate.
    ▪ Fast and convenient: single reagent “mix-and-measure” assay allows quantitation of free phosphate within 30 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 plates. Can be readily automated on HTS liquid handling systems
    
    Please contact us at for specific academic pricing.
- Properties
  - Details
    
    Method: OD620nm (malachite green)
    Samples: Enzyme reactions with free phosphate releases
    Species: All
    Procedure: 30 min
    Detection Limit: 0.4 µM
    Shelf Life: 12 months
    
    Storage
    
    4°C
    
    * For Research Use Only
- Applications
  - Application Description
    
    For sensitive and high-throughput phosphate determination.
- Reference
  - Christensen, T., et al. (2020). In vitro characterization of a novel consensus bacterial 6-phytase and one of its variants. Current Biochemical Engineering 6(3): 156-171. Assay: Phosphate in Trichoderma reesei phytase.
    Nitta, K., et al. (2021). Investigation of the effects of actinorhodin biosynthetic gene cluster expression and a rpoB point mutation on the metabolome of Streptomyces coelicolor M1146. Journal of Bioscience and Bioengineering. Assay: Phosphate in Streptomyces coelicolor M1146 cells.
    Nitta, K., et al. (2020). Multi-omics analysis of the effect of cAMP on actinorhodin production in Streptomyces coelicolor. Frontiers in Bioengineering and Biotechnology 8: 595552. Assay: Phosphate in bacteria nutrition medium.
    Lejeune, C., et al. (2021). Impact of phosphate availability on membrane lipid content of the model strains, Streptomyces lividans and Streptomyces coelicolor. Frontiers in Microbiology 12: 623919. Assay: Phosphate in R2YE agar medium.
    Hwang, J. D., Ortiz-Maldonado, M., & Paramonov, S. (2016). Delivery of Formulated Industrial Enzymes with Acoustic Technology. Journal of laboratory automation, 21(1), 153-165. Assay: Phosphate in Buttiauxella sp Phytase.
    Demidenko AA et al (2011). Effects of viscogens on RNA transcription inside reovirus particles. J Biol Chem. 286(34):29521-30. Assay: Phosphate in rat lipid.
    Liu Q 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 mouse intracellular phosphate.
    Lopez-Vales R, et al (2010). Fenretinide promotes functional recovery and tissue protection after spinal cord contusion injury in mice. J Neurosci. 30(9):3220-6. Assay: Phosphate in mouse lipid.
    Oborna I et al (2010). Increased lipid peroxidation and abnormal fatty acid profiles in seminal and blood plasma of normozoospermic males from infertile couples. Hum Reprod. 25(2):308-16. Assay: Phosphate in fish marine teleost follicles, eggs.
    Oborna I et al. (2010). Increased lipid peroxidation and abnormal fatty acid profiles in seminal and blood plasma of normozoospermic males from infertile couples. Hum Reprod. 25(2):308-16. Assay: Phosphate in human blood, semen.
    Polewski MD, et al (2010). Inorganic pyrophosphatase induces type I collagen in osteoblasts. Bone 46(1):81-90. Assay: Phosphate in mouse/human cellular extract.
    Cramp RL et al (2009). Ups and downs of intestinal function with prolonged fasting during aestivation in the burrowing frog, Cyclorana alboguttata. J Exp Biol. 212(22):3656-63. Assay: Phosphate in mouse lipid.
    Cramp RL, et al (2009). Ups and downs of intestinal function with prolonged fasting during aestivation in the burrowing frog, Cyclorana alboguttata. J Exp Biol. 212(22):3656-63. Assay: Phosphate in frog Na+/K+-ATPase extract.
    Huxtable AG, et al (2009). Tripartite purinergic modulation of central respiratory networks during perinatal development: the influence of ATP, ectonucleotidases, and ATP metabolites. JNeurosci. 29(47):14713-25. Assay: Phosphate in rat ATPase extract.
    Kang HY et al (2008). Altered TNSALP expression and phosphate regulation contribute to reduced mineralization in mice lacking androgen receptor. Mol Cell Biol.28(24):7354-67. Assay: Phosphate in rat ATPase extract.
    Kang HY, et al (2008). Altered TNSALP expression and phosphate regulation contribute to reduced mineralization in mice lacking androgen receptor. Mol Cell Biol.28(24):7354-67. Assay: Phosphate in mouse cytosolic fraction.
    Chen X et al (2007). The isolation and structure of membrane lipid rafts from rat brain. Biochimie 89(2):192-6. Assay: Phosphate in mouse/human cellular extract.
    Chen X, et al (2007). The isolation and structure of membrane lipid rafts from rat brain. Biochimie 89(2):192-6. Assay: Phosphate in rat lipid.
    Hough TA et al (2007). Novel mouse model of autosomal semidominant adult ypophosphatasia has a splice site mutation in the tissue nonspecific alkaline phosphatase gene Akp2. J Bone Miner Res. 22(9):1397-407. Assay: Phosphate in frog Na+/K+-ATPase extract.
    Hough TA, et al (2007). Novel mouse model of autosomal semidominant adult hypophosphatasia has a splice site mutation in the tissue nonspecific alkaline phosphatase gene Akp2. J Bone Miner Res. 22(9):1397-407. Assay: Phosphate in mouse intracellular phosphate.
    Lau K.H. et al. (2006). An osteoclastic protein-tyrosine phosphatase is a potential positive regulator of the c-Src protein-tyrosine kinase activity: a mediator of osteoclast activity. J Cell Biochem. 97(5):940-55. Assay: Phosphate in rabbit phosphatases.

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PiBlue™ Phosphate Assay Kit

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