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Fluoro Phosphate

Fluoro Phosphate Cell Technology Metabolic Assays
Fluoro Phosphate
Fluorescent Phosphate Detection Kit

$225.00

in stock
Product code: FLPHOS100
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Product Description

Introduction

Phosphorus is essential for multiple and diverse biological functions, including cellular signal transduction, mineral meatbolism, and energy exchange. Elevated levels of serum phosphate may be associated with a higher risk of death and adverse cardiovascular outcomes in people with prior myocardial infarction, and is the subject of large amount of research.

Key Benefits

  • Quantitation of phosphate anion in blood, plasma and serum samples.
  • Sensitive – can detect upto 200nM phosphate
  • Easy to use – Fluorescence (ex/em 535/585nm or Absorbance readout @ 570nm)

Assay Principle

Cell Technology’s Phosphate detection kit provides a simple, one-step fluorimetric or colorimetric method for determination of phosphate in serum and plasma samples. The assay is based on an enzyme-coupled reaction that detects inorganic phosphate. The substrate is converted to Hydrogen peroxide in presence of inorganic phosphate in a coupled enzymatic reaction. The Hydrogen Peroxide then reacts with the detection reagent in a 1:1 stoichiometry in presence of Horse Radish Peroxidase to produce the stable fluorescent product.

Substrate + Pi

↓ Coupled enzyme reaction

H2O2

↓ Horse Radish Peroxidase + Detection Reagent

Fluorescent Product

λmax 570nm. λ ex/em 535/585nm.

Colorimetric assay can be read on a spectrophotometer at 570nm.

Fluorescence is measured at excitation 530nm and emission 585nm.

Fig.1 Phosphate standard curve was generated using fluorimetric detection: excitation 535nm and emission 585nm. Incubation time= 60 minutes at 37oC. Standard curve range 1.5625 µM to 100µM. Graph was plotted using 4PL non-linear regression.

Fig.2 Phosphate standard curve inset: R2 =0.9835. Incubation time=60 minutes at 37oC. Standard curve range 1.5625µM to 25µM. Graph plotted with Linear regression.

References

  • Mohanty, J.G. et al : A highly sensitive fluorescent micro-assay of H2O2 release from activated human leukocytes using a dihydroxyphenoxazine derivative, J.Immunological Methods, 202, issue 2, P.133-141 (1997)
  • Tonelli, M. et al : Relation between serum phosphate level and cardiovascular event rate in people with coronary disease, Circulation,112, P.2627-2633 (2005)

Kit contents and Long Term storage

ReagentPart#Temperature
25X SubstratePart # 7023-20C
Detection ReagentPart # 4025-20C
Enzyme APart # 6026-20C
Enzyme BPart # 6027-20C
Phosphate StandardPart # 30602-8C
Horseradish Peroxidase EnzymePart # 60282-8C
1X Reaction BufferPart # 30592-8C
Sample DiluentPart # 30612-8C
1 Black Clear-Bottom 96-Well Plate for Fluorescent Plate Reader
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