Cell Signaling Technology

Product Pathways - Phosphatases

Malachite Green Phosphate Detection Kit #12776

REACTIVITY

No. Size Price
12776S 1 Kit ( 500 assays ) ¥1,672.00 现货查询 购买询价
Kit Includes Quantity Applications Reactivity Homology† MW (kDa) Isotype
Malachite Green Reagent 28 ml
Phosphate Standard (1 mM) 0.5 ml

Specificity / Sensitivity

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.

Description

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.

Phosphatase Assay

Phosphatase Assay

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.

Phosphatase Assay

Phosphatase Assay

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).

Background

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).

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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