Many people face challenges when using ELISA kits (Enzyme-Linked Immunosorbent Assay). Whether it's an issue with the standard curve, recovery rate, or other experimental problems, Shanghai Jinma is here to help you overcome these difficulties and achieve accurate results.
During your experiment, you might encounter several common issues. Here are some practical solutions:
Curve Problems:
- Abnormal OD values: Make sure the kit has been properly warmed up to 25°C. Check incubation temperature and time accuracy. Avoid excessive shaking or prolonged soaking during washing. Wash 4–5 times with 250 µL per well. Dry the plate in direct sunlight or avoid strong air currents. Ensure the microplate reader is set to the correct wavelength (usually 450 nm).
- White background or no signal: Incorrect preparation of washing solution can cause this. Always follow the instructions carefully—add the required amount of liquid, don’t skip any steps.
- No gradient on the standard curve: Contamination of the standard solution may be the cause. Check that the OD values of the standard points are within normal range. Ensure proper temperature control and avoid manual errors when setting up the plate.
- Poor linearity: Check if the OD values are consistent. If the curve is not linear, recheck the temperature and incubation conditions. Also, ensure that the sample and reagents are prepared correctly.
Recovery Rate Issues:
- High blank value: The blank tube may have been contaminated. Replace it with a fresh one. If the sample itself is positive, use a negative control instead.
- High recovery rate: Inaccurate pipetting can lead to this problem. Use precise volume measurements and avoid adding too much or too little reagent. Dry the solution thoroughly and avoid cross-contamination between samples.
- Low recovery rate: This could be due to inaccurate dilution or incorrect addition of the sample. Double-check the dilution process and ensure the complex solution is properly prepared. A false negative result may also occur due to water quality issues.
Other Common Causes:
- Water quality: Always use high-quality water, such as Wahaha mineral water, for preparing solutions.
- Experimental operation: Ensure thorough drying of all phases and avoid contamination during centrifugation. If the sample is emulsified, try a warm bath at 70°C to resolve the issue.
- Instrument or reagent issues: Clean all equipment thoroughly before use. Check if the reagent blank shows any interference. Organic solvents may affect the results, so use them cautiously.
- Derivatization reagent degradation: If the reagent has been stored for a long time, it may lose effectiveness. Replace it if necessary.
We provide free testing services with our ELISA kits. You only pay for the kit, while all other reagents and support are included at no extra cost. Based on your experimental workload and difficulty level, we will agree on a timeline to complete your experiment with high quality and deliver the results and data to you for free. Contact us today and experience reliable, accurate, and hassle-free ELISA testing!
Mitsubishi Truck Starter Parts
truck starter，Mitsubishi Truck Starter Parts， truck Starter Parts，
YIWU JINGHONG AUTO PARTS CO.,LTD , https://en.jhauto.ru