Isolation of Human PBMC from Whole Blood
Cryopreservation of Human Plasma
1. Principle
1. Samples may be collected in Heparin, sodium citrate or EDTA tubes. Use care when using heparin as an anticoagulant since an excess of heparin will give falsely high values. Use no more than 10 IU of heparin per mL of blood collected.
2. Human Plasma can be frozen at -80(C and tested later.
2. Materials and Equipment
1. Patient sample in Green-top (Heparin), Purple-top (EDTA), or Light Blue-top (sodium citrate) tubes.
2. Freezing box
3. -80(C freezer
4. Biosafety cabinet
5. Centrifuge
3. Procedure
1. Centrifuge samples at 500 RCF at (RT) Room Temperature for 10 minutes (cells in pellet).
2. Collect the supernatant and transfer into a clean tube.
3. Centrifuge supernatant (from step 3.2) at 13,000 RCF in a table-top microfuge at room temperature for 10 minutes.
4. Aliquot and use immediately or store at –80°C.
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