In ELISA experimental operation, sample addition is an important step, and this step has a great influence in the whole experiment. To organize this issue.
- the possible causes of the spiking problem
1. poor separation of serum or plasma specimens that are spiked.
2. manual operation, adding too much sample plate caused by adding samples into the incubator before waiting too long (especially when the room temperature is high).
3. Enzyme splashing out of the well when adding enzyme reagent after specimen addition
1. specimen is serum: blood will be stored naturally for 1-2 hours first, then centrifuged at 3000rmp for 15 minutes.
Specimen as plasma: blood specimen collection tubes containing anticoagulant must be used, and blood collection tubes must be mixed upside down 5-10 times immediately after collection, placed for a period of time, and then centrifuged at 3000rpm for 15 minutes; if tested within a few days, it can be placed in a refrigerator at 2-8℃, or in a low temperature refrigerator at -20℃ if it is to be stored.
2. Put into incubator promptly after adding samples.
3. After adding the enzyme reagent, wipe the surface of the enzyme plate lightly with absorbent paper to absorb it.
4. If AT or other fully automatic spiking is used, choose FAME or other post-processing instruments to add enzyme reagents.
5. When there are more specimens, please operate in batches.