Quantitative EIA for the determination of 25-hydroxyvitamin D (25-OH D) and other hydroxylated metabolites in human serum or plasma.
This method calculates the percentage binding B/B0% of each sample from the zero calibrator (Standard1). A calibration curve is prepared on semi-log axes plotting B/B0% against concentration values with a Four Parameter Logistic curve (4PL) fitted through the data. The the nmol/L is calculated for each sample with specified dilution factor values applied.
Supply your absorbance data as reference corrected (i.e. measured at 450nm with reference measured at 650nm).
If the entire plate was read then supply the measured value for each well. Alternatively, if only specific parts of the plate were read then list only those measured values. In this case an additional step is required: under the Microplate section below, select or create a layout which defines which wells were read and which were not. More...
A zero concentration value has been specified with a logarithmic x axis.
This is valid but means that you will not see the zero standard on the chart, also in certain cases it may result in a skewed fit. One alternative is to use a suitably low value, such as 0.01 instead of zero. More...
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