Specimen collection and handling for ELISA
Specimens should be clear, non-hemolyzed and non-lipemic. Excessive hemolysis and the presence of large clots or microbial growth in the sample may interfere with the performance of the test.
- Cell culture supernatant: remove any particulate matter by centrifugation.
- Serum: use a clot tube and allow sample to clot for 30-45 min at RT, then centrifuge for 10-15 min at 1,000-2,000 x g (RT) and collect serum immediately.
- Plasma: collect plasma by using anticoagulant, such as EDTA or heparin. Mix well immediately after collection. Centrifuge for 10-15 min at 1,000-2,000 x g (RT) and collect plasma.
Samples should be aliquoted and stored frozen at ≤-20 °C to prevent cytokine degradation. If samples are run within 24 hours, they may be stored at 2-8 °C. Avoid repeated freeze-thaw cycles. Do not heat serum or plasma samples. Prior to assay, frozen samples should be completely thawed and mixed well.
Note:
Specimen collection from humans and non-human primates should be carried out in accordance with CLSI document M29-A4 “Protection of laboratory workers from occupationally acquired infections”. No known test method can offer complete assurance that human- or non-human primate-derived blood samples will not transmit infection. Therefore, all human and non-human primate specimens should be considered potentially infectious.
RT: room temperature (temperature between 20 °C and 26 °C)