The accuracy of immunodetection shows the repeatability between wells (within batches) in a single experiment and between multiple experiments (between batches). A high CV value indicates low accuracy, which is usually caused by two reasons: pipetting technology and washing technology.
Perceiving technology
1. When pipetting, the gun head should be aimed at the center of the hole to avoid contacting the wall and bottom of the hole.
2. Shake the mixture gently after pipetting.
3. If your sample is sticky, please wash the gun head in advance to eliminate the effect of surface tension. Drink the liquid and wait for a while to equilibrate the volume before removing it.
4. Inadequate pipetting or uneven fluid accumulation indicates that the pipette needs to be corrected. Check the pipette and reset it if necessary.
5. When adding samples, different reagent standards and samples should ensure that the gun head is replaced to avoid cross-contamination. Make sure to use the right head of the gun. The inappropriate gun head cannot properly measure the volume of aspirate and sample.
Washing technology
1. If using an automatic plate washing machine or multi-channel pipette, make sure the inlet and exit operate normally.
2. After the last wash, flip the microplate to pour out the excess wash liquid and pat dry the liquid on the absorbent paper. Washing the plate too fast or too slow reduces the accuracy. The liquid in the hole cannot be dried naturally, and the next operation should be performed immediately.
3. Add a sufficient amount of washing liquid to the well according to the instructions, and ensure that the washing is complete.
4. Since the washing solution is not universal, when you use up the washing solution in the kit, please do not use the washing solution in other kits. If necessary, contact technical support for assistance.
5. Do not reduce or increase the soaking time or skip the soaking step.
6. Wash according to the number of washing times recommended in the instructions, and changing the number of washing times arbitrarily will affect the accuracy of the experiment.
Storage liquid bottle
To avoid contamination, different liquid storage bottles were used for different reagents. Some reagents or samples are easily contaminated (such as saliva, oxidants, etc.), please refer to the instructions. Take some precautions to protect the solution when necessary. In addition, to ensure the cleanliness of the reagent or detergent sample tank, to avoid causing pollution.
Cover the plating film
Used sealing plate cladding may contain reagents remaining from the previous experiment, which can cause well plate contamination if repeated, resulting in low accuracy. A new sealing plate coating is recommended for each incubation.