Hu Ding shares with you how to draw the standard curve of ELISA kits - Database & Sql Blog Articles

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Drawing a standard curve is one of the essential procedures in the elisa kit testing process. The numerous operational steps of the ELISA kit determine its many influencing factors.

Therefore, in the quantitative determination, each batch of tests must be prepared with a series of different concentrations of reference standards under the same conditions. Hu Ding shares with you how to draw a standard curve for an ELISA kit.

Various mapping software can be used to draw the ELISA standard curve. Our company takes "Curve Exert1.3" software as an example, and the ELISA standard curve is as follows:


Taking the concentration of the standard as the abscissa and the OD as the ordinate, draw a standard curve on the coordinate paper, and find the corresponding concentration from the standard curve according to the OD value of the sample; multiply by the dilution factor; or use the concentration of the standard Calculate the linear regression equation of the standard curve with the OD value, substitute the OD value of the sample into the equation, calculate the sample concentration, and multiply by the dilution factor, which is the actual concentration of the sample.

There are several issues to be aware of when doing standard curve sample testing.

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1. The concentration of the sample and other indicators are calculated according to the standard curve, so it is important to consider the standard curve as a more important thing than the formal experiment. Otherwise, the results of the latter experiment cannot be discussed.

2. Set the standard concentration range of the standard curve sample to have a relatively large span, and to cover the concentration of the test sample you want to test, that is, the concentration of the sample should be within the concentration range of the standard curve, and the human ELISA kit includes the upper limit and Lower limit. For the standard curve of S type, try to make the concentration of the experimental sample in the steepest section of the middle slope, that is, the curve is almost in a straight line.

3. It is best to prepare the standard sample concentration in the standard curve by the double dilution method. The ELISA kit can ensure that the concentration of the standard sample does not deviate greatly.

4. When testing standard samples, they should be carried out in increasing order of concentration to reduce the influence of high concentration on low concentration and improve accuracy.

5. The number of samples in the standard curve is generally 7 points, but at least 5 points must be guaranteed.

6. The correlation coefficient of the standard curve made varies according to the experimental requirements, but in general, the correlation coefficient R is at least greater than 0.98. For some experiments, at least 0.99 or even 0.999.

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