GENTAUR - Research solutions for the academic, biotechnology and pharmaceutical industries
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Fluoro Phosphate NEW!
Fluorescent Phosphate Detection Kit
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| 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)
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| 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. |
| Assay Principle: |
Gentaur’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.
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| Kit contents (for 100 assays) |
| 1. |
25X Substrate ......................................................................Part # 7023 -20°C |
| 2. |
Detection Reagent ................................................................Part # 4025 -20°C |
| 3. |
Enzyme A ...........................................................................Part # 6026 -20°C |
| 4. |
Enzyme B ...........................................................................Part # 6027 -20°C |
| 5. |
Phosphate Standard ..............................................................Part # 3060 2-8°C |
| 6. |
Horseradish Peroxidase enzyme ...............................................Part # 6028 2-8°C |
| 7. |
1X Reaction Buffer ...............................................................Part # 3059 2-8°C |
| 8. |
Sample Diluent ...................................................................Part # 3061 2-8°C |
| 9. |
1 Black clear-bottom 96-well plate for fluorescent plate reader |
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