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12776
Malachite Green Phosphate Detection Kit
Cellular Assay Kits

Assay Kit - Malachite Green Phosphate Detection Kit - 1 Kit #12776

Phosphatase Assay Image 1 - Malachite Green Phosphate Detection Kit

Figure 1. A linear standard curve was generated using phosphate standards and the Malachite Green Phosphate Detection Kit. The phosphate standard was diluted in dH2O as described in the protocol section and samples were assayed with the Malachite Green Phosphate Detection Kit. This standard curve is for demonstration purposes only; users should generate a standard curve for each sample set in order to accurately determine phosphate concentration.

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Phosphatase Assay Image 2 - Malachite Green Phosphate Detection Kit

Figure 2. Receptor type protein phosphatase F (PTPRF) activity was measured using the Malachite Green Phosphate Detection Kit. PTPRF phosphatase activity was monitored using tyrosine phosphopeptide-2 (DADE(pY)LIPQQG) as the substrate. The reactions were performed with various amounts of PTPRF phosphatase at 37ºC for 10 minutes in a buffer containing 50 mM imidazole, 0.2% 2-mercaptoethanol, 65 ng/μl BSA, and 1 mM tyrosine phosphopeptide-2 (DADE(pY)LIPQQG).

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Product Includes Quantity (with Count) Storage Temp
Malachite Green Reagent 2 x 28 ml RT
Phosphate Standard (1 mM) 1 x 0.5 ml RT

The Malachite Green Phosphate Detection Kit is a convenient and sensitive, single-step free-phosphate determination kit that can be used for measuring phosphate released during enzymatic phosphatase assays.

The Malachite Green Phosphate Detection Kit detects only free-phosphate in sample. Figure 1 demonstrates a linear range of 50 to 1600 pmol of phosphate.

The Malachite Green Phosphate Detection Kit is designed for the in vitro measurement of phosphatase activity. Protein phosphatase enzymatic activity results in the release into solution of free inorganic phosphate from protein-phosphatase substrates. This free inorganic phosphate then forms a green molybdophosphoric acid complex. Formation of this complex is monitored and quantified by measuring the absorbance at 630 nm. This absorbance is proportional to free-inorganic phosphate in solution and correlates with the activity of phosphatase (1,2).

  1. Baykov, A.A. et al. (1988) Anal Biochem 171, 266-70.
  2. Geladopoulos, T.P. et al. (1991) Anal Biochem 192, 112-6.
For Research Use Only. Not For Use In Diagnostic Procedures.

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