The details make the difference, and this is especially true for laboratory researchers. In experiments, even small mistakes can lead to failure, which is why attention to detail is crucial. In this technical article, Shanghai Jinma Laboratory shares key insights on analyzing ELISA kit results.
ELISA (Enzyme-Linked Immunosorbent Assay) is a widely used technique in biological research. However, the success of an ELISA experiment depends heavily on proper execution and accurate data interpretation. One of the most important steps is ensuring that samples are tested in duplicate or triplicate, and the average OD value is calculated for each standard concentration. The coefficient of variation (CV) should be less than 20% to ensure reliability.
After obtaining the OD values, the next step is to subtract the blank well's OD from the sample readings. This helps eliminate background noise and provides more accurate results. Then, a standard curve is plotted by placing the corrected OD values on the X-axis and the known concentrations on the Y-axis.
To create the best fit curve, software like CurveExpert 1.4 is recommended. It offers multiple fitting methods—such as linear, semi-log, log, and four-parameter logistic models. You can test different equations to see which one gives the closest result when you input the OD values into the equation. The ideal result should be within ±10% of the actual concentration.
If the standard curve doesn't show good linearity, or if there's poor parallelism between replicate wells, it could be due to contamination, improper washing, or incorrect pipetting techniques. To avoid these issues, always handle reagents carefully, avoid splashing during washing, and use a calibrated micropipette. Make sure not to touch the tip of the pipette to the well, and replace the tip after each transfer. Also, keep the plate covered and in a stable incubation environment to prevent evaporation.
Shanghai Jinma Lab uses high-quality imported materials and offers free testing, data analysis, and technical support. Their products are trusted by top hospitals, research institutes, and universities across the country. With years of experience and a commitment to excellence, they have earned widespread recognition in the scientific community. If you're looking for reliable ELISA kits and expert guidance, contact Shanghai Jinma today.
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