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MicroMolar Iodide Assay Kit

BHE16300285

This product includes 40 ul of 100 x Reagent A, 40 ul of 100 x Reagent B, 200 ul of 100 x fluorescence dye and 50 ul of 10 mM potassium iodide solution.

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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 150 µl and the final assay volume is 290 µl. Please adjust the sample volume and reagent volume proportionally if a 384-well plate or a cuvette is used for the measurement.
 
STANDARD CURVE
1. Sample preparation: Prepare 150 µl of potassium iodide solutions in a 96-well black plate with a two-fold serial dilution from 10 µM to zero in water or a 10 mM HEPES, pH 7.4 buffer. Freshly make the 1 x reagents by 10-fold dilution of the 100 x Reagents with water.
 
2. Detection: Mix 20 µl of 1 x Reagent A with 150 µl of the iodide solutions and incubate the mixture for 5 min. Then add 20 µl of 1 x Reagent B and mix the solution and incubate the mixture for 5 min. Finally mix 100 µl of 1 x fluorescence dye with the sample and read the fluorescence at 535 nm with excitation at 485 nm in 5 min.
 
                Note: a longer incubation after addition of the dye will yield a higher assay sensitivity but compromise the assay linearity.
 
3. Data Analysis: Plot the fluorescence intensity Fc and the cysteine concentration [Iodide] to generate the linear standard curve.
 
                                                                  Fc = a [Iodide] + 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 Iodide concentrations.
 
UNKNOWN SAMPLES
Follow the same procedure to measure the fluorescence intensity Fc values from the unknown samples. Calculate the Iodide concentrations in the unknown samples using the Fc values from the unknown samples and the a and b values from the standard curve.
 
                                                                [Iodide] = (Fc – b) / a
SKU Supplier No. Size Your price Quantity
BHE16300285 MIA200 200 assays $289.02
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