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Home / Assay Kits

MicroGram Vancomycin Assay Kit

BHE16300276

This product includes 0.5 ml of 10 x VAN dye, 5.5 ml of Assay buffer, 5 ml of reagent A. It is for measurement of 100 samples using 96-well plates.

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Product Manual Datasheet
Protocols Description ASSAY PROTOCOL
The following assay protocol is based on assays using a 96-well plate for the measurement. The sample volume is 50 µl and the final assay volume is 100 µl. For 384-well plate assays, the sample volume is 30 µl and the final assay volume is 60 µl. For assays using cuvette, the sample volume is 500 µl and the final assay volume is 1000 µl.
 
STANDARD CURVE
1. Sample preparation: Prepare 50 µl of vancomycin in the wells of a black 96-well plate with a two-fold serial dilution from 100 µM to zero in Assay Buffer. For 10 samples, dilute 0.05 ml of the 10 x VAN dye 10-fold with 0.45 ml of Reagent A to make 0.5 ml of 1 x VAN dye.
 
2. Detection: Mix 50 µl of 1 x VAN dye with 50 µl of the vancomycin solutions for 5 min and read the fluorescence at 470 nm (excitation at 390 nm).
 
3. Data Analysis: Plot the fluorescence intensity Fc and the vancomycin concentration [Vancomycin] to generate the linear standard curve.
 
                                                                  Fc = a [Vancomycin] + 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 vancomycin concentrations.
 
UNKNOWN SAMPLES
Follow the same procedure to measure the fluorescence intensity Fc values from the unknown samples. Calculate the vancomycin concentrations in the unknown samples using the Fc values from the unknown samples and the a and b values from the standard curve.
 
                                                                 [Vancomycin] = (Fc – b) / a
SKU Supplier No. Size Your price Quantity
BHE16300276 VAN100 100 assays $448.76
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