The details make all the difference, and this is especially true for laboratory researchers. In today's technical article, Shanghai Jinma Lab brings you a detailed guide on how to analyze ELISA kit test results effectively.
In ELISA experiments, it's crucial to run samples in duplicates or triplicates, and then calculate the average OD value for each concentration standard. The coefficient of variation (CV) between duplicate wells should be less than 20% to ensure reliable data.
Next, subtract the OD value of the blank well from the average OD value of each sample. This step helps eliminate background noise and improves accuracy.
After that, create a standard curve by plotting the adjusted OD values on the X-axis and the corresponding standard concentrations on the Y-axis. Use software like CurveExpert 1.4 to generate the most suitable fitting equation—whether linear, semi-logarithmic, logarithmic, or four-parameter logistic. Test the accuracy by plugging in known concentrations into the equation and checking if the calculated values are within ±10% of the actual values.
If the standard curve shows poor linearity or if there's significant variation between duplicate wells, it may be due to contamination between wells, improper washing, or incorrect pipetting techniques. To avoid these issues, make sure not to spill liquid into other wells when adding samples or reagents. When washing manually, avoid splashing the wash buffer into adjacent wells. Always use a new pipette tip when transferring liquids from different bottles and ensure the incubation environment is dark and the plate is properly sealed with a cover film.
Additionally, use a calibrated micropipette and always drip the last drop onto the well to maintain consistent volume. These small but important steps can significantly improve the reliability and reproducibility of your ELISA results.
Shanghai Jinma Lab uses high-quality imported materials and offers free testing, data analysis, and technical support. Our products are widely used across hospitals, research institutes, and academic institutions nationwide. With a strong commitment to quality and scientific excellence, we have earned widespread recognition in the field of bioscience. Contact us today to learn more about our products and services.
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