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MicroMolar Polyphosphate Assay Kit

BHE16300259

This product includes 300 ul of 100 x PPD dye and 30 ul of 1 mM sodium polyphosphate ( a 45-mer). It is for 1000 assays using 384-well plates (30 ul of sample volume) or 500 samples using 96-well plates (60 ?l of sample volume). Cuvettes may also be used for measurements.

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+1 866.986.9598
Email
[email protected]
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Reference 1. Achbergerová L. et al, Degradation of polyphosphates by polyphosphate kinases from Ruegeria pomeroyi. Biotechnology Letters. Volume 36, Issue 10, pp 2029-2035 (2014).
2. Zhang J. et al, A Fast Sensor For in Vivo Quantification of Cytosolic Phosphate in Saccharomyces Cerevisiae, Biotechnol Bioeng.Vol 112, pp 1033-1046 (2015).  
Product Manual Datasheet
Protocols Description PROTOCOL
Standard curve
1. Sample preparation: Prepare the polyphosphate solutions, 45-mer sodium polyphosphate, at a series of concentrations ranging from 50 µM to zero in a 10 mM HEPES, pH 7.4 buffer. Dilute the 100 x PPD dye 100-fold with water to make the 1 x PPD dye.
 
Note: The fluorescence intensity has a linear relationship with the polyphosphate concentrations in the range of 1 to 50 µM. A polyphosphate concentration higher than 50 µM suppresses the fluorescence signal.
 
2. Detection: Mix 30 µl of the sample with 30 µl of the 1x PPD dye solution for 5 min. Read the fluorescence intensity at 550 nm (excitation 415 nm).
 
3. Data Analysis: Plot the fluorescence intensity Fc and the polyphosphate concentration [Polyphosphate] to generate the linear standard curve.
 
                                                                                  Fc = a [Polyphosphate] + b
 
Where the Fc values are from experimental data, the a and b values are from the linear fitting between the Fc values and the polyphosphate concentrations.
 
UNKNOWN SAMPLES
 
 Follow the same procedure to measure the fluorescence intensity Fc values from the unknown samples. Calculate the polyphosphate concentrations in the unknown samples using the Fc values from the unknown samples and the a and b values from the standard curve.
 
                                                                                [Polyphosphate] = (Fc – b) / a
SKU Supplier No. Size Your price Quantity
BHE16300259 PPD1000 1000 assays $358.75
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