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MicroMolar EDTA Assay kit

BHE16300284

This product includes 200 ul of 100 x C56 dye and 1000 ul of 1 mM EDTA. It is for measurement of 200 samples using 96-well plates. Cuvettes may also be used for measurements.

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[email protected]
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Product Manual Datasheet
Protocols Description ASSAY PROTOCOL
The following assay protocol is based on using a 96-well plate for the measurement. The sample volume is 100 µl and the final assay volume is 200 µl. For 384-well plate assays, the sample volume is 40 µl and the final assay volume is 80 µl. For assays using cuvette, the sample volume is 500 µl and the final assay volume is 1000 µl.
 
STANDARD CURVE
1. Dye dilution: For 10 samples, dilute 0.011 ml of the 100 x C56 dye 100-fold with water to make 1.1 ml of 1 x C56 dye. Keep the solution at room temperature for 5 min.
 
2. Sample preparation: Prepare 100 µl of EDTA solutions in the wells of a black 96-well plate with a two-fold serial dilution from 0.1 mM to zero in 10 mM HEPES, 0.1 M NaCl, pH 7.4 buffer.
 
3. Detection: Mix 100 µl of 1 x dye C56 with 100 µl of the EDTA solutions for 5 min and read the fluorescence at 535 nm (excitation at 485 nm).
 
Note: The assay sensitivity is higher when the incubation time after dye addition is longer. The assay linearity will be also affected by the incubation time.
 
4. Data Analysis: Plot the fluorescence intensity Fc and the EDTA concentration [EDTA] to generate the linear standard curve.
 
                                                                         Fc = a [EDTA] + 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 EDTA concentrations.
 
UNKNOWN SAMPLES
Follow the same procedure to measure the fluorescence intensity Fc values from the unknown samples. Calculate the EDTA concentrations in the unknown samples using the Fc values from the unknown samples and the a and b values from the standard curve.
 
                                                                         [EDTA] = (Fc – b) / a
 
SKU Supplier No. Size Your price Quantity
BHE16300284 EDTA200 200 assays $302.17
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