How to Draw a Standard Curve for ELISA?
Several software programs can be used to create an ELISA standard curve. One commonly used tool is "CurveExpert 1.3." Below is a step-by-step guide on how to draw the ELISA standard curve using this software.
1. Launch "CurveExpert 1.3" on your computer.
2. Input the standard OD values on the X-axis and the corresponding concentrations on the Y-axis. A sample graph will appear, as shown in the interface.
3. Click the [Run] button. A dialog box will pop up. Select [OK] to proceed. Two additional windows may appear; close the second one if it appears.
4. In the upper right corner of the main window, several curve options are listed. Click on the curve name labeled "1" to view the corresponding fit in the lower right section.
5. Choose the curve that best fits your ELISA data. Double-click the selected curve to open the fitting details.
6. Note: To ensure the best fit, check the coefficient "R" value. The closer the R value is to 1, the better the curve fits the data.
7. Press [Ctrl] + [L] to open the calculation window.
8. Enter the measured OD value of the unknown sample and click [Calculate]. This will give you the actual protein concentration. If the sample was diluted N times, multiply the result by N to get the true concentration.
9. To obtain the equation of the fitted curve, right-click on the blank area in the graph window and select “Information.â€
10. A new dialog box will appear with the curve equation. Copy the relevant data as needed. For example:
**RationalFunction:** y = (a + bx) / (1 + cx + dx²)
**CoefficientData:** a = -21.531017, b = 212.05019, c = -0.24054479, d = 0.018060778
This process ensures accurate quantification of protein levels in ELISA experiments. Make sure to validate your results with appropriate controls and repeat measurements when necessary.
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