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Isolating NHP Serum and Plasma

Created by Ravishankar Madhu

Last updated: 2023-07-10

Description

This protocol will describe how to process and isolate NHP Serum and Plasma.

Reagents and Materials

  • Non Human Primate Peripheral Blood
  • Non Human Primate SST Tube with blood

Things to Note before starting

Estimated Timing

The process involves a 15 minute spin that occupies the majority of this protocols time. Note that the spin speed is abnormal, so it is unlikely the centrifuge can be shared with others while performing this spin.

Procedure

Isolating Plasma from PB

  1. Aliquot 1mL of NHP PB into a FACS tube. 1
  2. Spin down FACS tubes at 2000 x g for 15 minutes
  3. After the spin, you should see a translucent solution on the top layer with darker red liquid on the bottom layer
  4. Using a p1000, carefully take up the translucent solution, making sure to not take up any blood, and aliquot into a 1.5mL Eppendorf tube.
  5. Label the tube with the Animal ID and date and store in the -80C Freezer.

Isolating Serum

  1. When you ask for Serum from the Primate Center, they will provide you with a Serum Collection tube. It contains a filter that allows blood to pass through at high speeds, but keeps the serum on the top layer.
  2. Balance the tube with a capped FACS tube and water and spin at 2000 x g for 15 minutes.
  3. After the spin, you should see a yellowish liquid above the filter. If their are tinges of red, spin again.
  4. Using a p1000, take up the yellowish liquid and aliquot into a 1.5mL Eppendorf tube.
  5. Label with the Animal ID and date and store in the -80C Freezer.

Next Steps

We do not really do a lot with the serum or plasma, but it is important to bank these samples for the lifetime of the project. During a NHP Transplant, baseline serum is collected and then used during infusion day to create a 2% autoserum solution

Appendix

Footnotes

  1. The more PB you add, the more plasma you will receive. I would estimate that you receive 40-50% of the input volume back as plasma. So on average 1mL of PB will give you 500uL of Plasma. ↩