Sample Collection & Storage

Serum

Use a red top separator tube (SST) and allow sample to clot for 30 minutes before centrifugation. Centrifuge for 15 minutes at approximately 1000g. Remove serum, aliquot, and store at -80°C.

Plasma

Collect plasma using EDTA, heparin, or citrate as an anticoagulant. Centrifuge for 15 minutes at approximately 1000g within 30 minutes. Remove plasma, aliquot, and store at -80°C.

Cell culture supernatents

Remove particulates by centrifugation, aliquot, and store samples at -80°C.

Urine

Collect the first urine of the day (mid-stream) in a sterile container. Centrifuge to remove particulate matter, aliquot, and store samples at -80°C.

Lysates

There is no defined protocol that we recommend for tissue extraction as this may differ for different tissue types and/or analyte one may wish to measure. Generally most protocols that are used in ELISAs can be used, but here are some guidelines in selecting a method.

1. Homogenize cells or tissues mechanically (eg. ultrasonication) in a PBS based buffer containing protease inhibitors (like aprotinin or an inhibitor cocktail) and low (< 0.2%) non-ionic detergent concentration.
2. Extraction medium should not contain any organic solvents like DMSO etc.
3. Centrifuge the extract, aliquot and store at -80°C.

CSF

Immediately after lumbar puncture, centrifuge the CSF at 800g for 5 minutes at 4°C. The CSF sample must be clear. A sample with blood contamination may compromise results. Aliquot the CSF samples and store at -80°C.

Isolight Sample Collection and Storage

For sample preparation, contact the Cytokine Reference Lab or refer to the following link from Isoplexis: https://isoplexis.com/support/sample-preparation/.