Serum and Plasma Matrices


Serum and Plasma are known for containing interference factors (hemolytic/bilirubin/lipemic/blood, etc) that may inhibit the assay's biological components (capture/detection antibodies), as well as potentially affect absorbance readings.


To mitigate suspected interference factors:

  1. Dilution is the best way to mitigate interference factors.  Dilute samples in the specified kit diluent as much as the target analyte concentrations will allow based on the dynamic range of the kit.
  2. Centrifuging samples at 10,000 rpm for 10 minutes at 4°C can remove some interference factors.
  3. Heterophilic or animal binding antibodies must be addressed with specific anti-interference buffers.


For ELISA, more dilution is always better to mitigate any potential assay interference. The optimal sample dilution level must be determined by serial dilution.  Where the sample measurements exhibit linear measurements from the back calculated linear regression from the standard curve across 2 or 3 dilutions is the optimal dilution point.


Plasma Anticoagulants


The best anticoagluant is dependent on the nature of the protein of interest. For all ELISA suppliers, we recommend contacting tech support if it is not explicitly stated. Most of Aviva's ELISA Kits are compatible with either EDTA, Citrate, and Heparin. We generally recommend using EDTA or Heparin if possible.


Sample Collection Methods

  • Serum - Use a serum separator tube (SST) and allow samples to clot for two hours at room temperature or overnight at 4°C before centrifugation for 15 minutes at 1,000 x g. Remove serum and assay immediately or aliquot and store samples at -20°C or -80°C. Avoid repeated freeze-thaw cycles.

  • Plasma - Collect plasma using EDTA, or heparin as an anticoagulant. Centrifuge for 15 minutes at 1,000 x g at 2-8°C within 30 minutes of collection. Assay immediately or aliquot and store samples at -20°C or -80°C. Avoid repeated freeze-thaw cycles.

Need more help? Send us an email at techsupport@avivasysbio.com.


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