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PPDK (Pyruvate, phosphate dikinase) Activity Assay Kit (Fluorometric/Colorimetric)

Pyruvate, phosphate dikinase, PPDK, (E.C. 2.7.9.1) is found in microorganisms and plants where it catalyzes reversible interconversion of AMP, PPi and phosphoenolpyruvate (PEP) to ATP, Pi and pyruvate monophosphate. PPDK plays an important role in plant photosynthesis and glycolytic pathway of endobiotic protozoa and bacteria where pyruvate, phosphate dikinase (PK) is absent; it functions in the process of ATP biosynthesis. In plants, PPDK is a pivotal regulator of the C4 photosynthetic cycle and the key limiting factor for maintaining photosynthesis at low temperature. It catalyzes the regeneration of PEP as the primary acceptor of atmospheric CO2 from pyruvate, ATP and phosphate. In addition, PPDK is also present in C3 leaves where it confers a similar light/dark regulation in chloroplasts, and in wheat leaves in an amount comparable to that found in seeds. Since PPDK is not found in vertebrates, finding its inhibitors may be useful for treatment of infections caused by anaerobic protists that depend on pyrophosphate-dependent glycolysis. Specific inhibition of PPDK is expected to inhibit Giardia and other anaerobic protists such as Entamoeba histolytica and Trichomonas vaginalis without harming their mammalian hosts. BioVision’s Pyruvate, phosphate dikinase, (PPDK) Assay Kit provides a simple and rapid test for measuring specific activity of PPDK in bacterial, protozoan and plant lysates as well as protein preps. We utilize the ability of PPDK to interconvert the substrate into intermediate product detected by proprietary enzyme mix and probe. Generated stable product can be quantified by either colorimetric or fluorometric readout. The assay is simple to perform, high-throughput adaptable and a fluorometric reaction can detect as low as of 0.006 U of PPDK activity in a single well. We provide sufficient reagents for 100 fluorometric or colorimetric assays.

Product Specifications

Reactivity

Bacterial/Plants

Applications

• Measurement of PPDK activity in tissue/cell lysates and protein preparations • Screening for specific inhibitors of PPDK activity that may affect microorganisms/plant biochemical pathways • Study of pyrophosphate-mediated glycolysis in bacterial and protozoan cell lysates • Detection of activity and light activation kinetics of PPDK from crude leaf extracts and recombinant protein • Study of photosynthesis in plants

Detection Method

Absorbance (570 nm) or Fluorescence (Ex/Em 535/587 nm)

Features & Benefits

• Simple procedure; takes ~ less than 60 minutes • Fast and convenient

Shipping Conditions

gel pack

Storage Conditions

-20°C

Shelf Life

12 months

Frequently Asked Questions

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