Before the incubation process begins, the sample loading time can be excessively long, which may affect the accuracy of the test. Additionally, when adding the enzyme reagent by dropping from a bottle, the angle and pressure applied during the drop can vary, leading to inconsistent volumes of reagent being added. This inconsistency can result in inaccurate test outcomes.
**Control Methods:**
1. When handling a large number of samples, it is advisable to process them in batches to minimize errors caused by prolonged experimental times.
2. After adding the enzyme reagent, use absorbent paper to gently remove any excess reagent on the surface of the wells.
3. For quantitative tests, always use a calibrated pipette or dispenser to ensure accurate reagent delivery. Regularly check and calibrate the pipettes to maintain precision. Also, perform proper controls to prevent cross-contamination between samples.
Incubation is one of the most critical and challenging steps in ELISA assays. The most commonly used incubation temperatures are 37°C and room temperature, with some kits also using 43°C or 2–8°C. In domestic ELISA kits, the typical incubation time at 37°C is 30 minutes to 1 hour, while imported kits often require 1 to 2 hours for a more complete reaction. The recommended incubation conditions should always follow the kit's instructions.
After adding the sample and/or reagent, the microplate is usually placed in a water bath or incubator at room temperature. However, it takes time for the temperature inside the wells to rise from room temperature to the desired incubation temperature (e.g., 37°C). If the timer starts as soon as the plate is placed in the incubator, the actual incubation time may be insufficient, especially for weak positive samples, leading to false-negative results.
**Control Methods:**
1. If multiple incubation temperatures are available, consider using a slightly lower temperature with a longer incubation time to improve consistency and sensitivity.
2. Place a small thermometer inside the microplate well along with the reaction solution to monitor the actual temperature during incubation.
3. Be aware of the "edge effect," where the outer wells may develop more intensely than the central ones due to temperature gradients across the plate. To reduce this, it is recommended to use a water bath instead of an incubator for more uniform heating.
By carefully controlling these factors, you can significantly improve the accuracy and reliability of your ELISA results.
BB Cream,Whitening Bb Cream,Bb Foundation Cream Tube,Moisturizing Foundation Bb Cream
RYEOGLOBAL Co,.LTD , https://www.ryeolab.com