Rapid separation and purification for PBMC within only 15 min.

Simple operation procedures: when collecting samples, directly decant the centrifuge tube. No other professionally technical operations are required. No need to slowly and laboriously add samples into the solution

Good reproducibility for reduced errors and less operation variation among users.

E-beam sterilized with SAL=106

No heat source; no nucleases

Preliminary Preparation

1) Equilibrate the separation solution away from light to the room temperature (RT) .

2) Prepare in advance the anticoagulated blood and bone marrow. It is suggested add saline to dilute the sample properly according to its density, so as to improve the separation performance.

1) Add the separation solution by a serological pipette through the central hole of the nested container: for a 15 ml tube, add approximately 4 ml separation solution; for a 50 ml tube, add approximately 13 ml separation solution; Please ensure the separation solution on top of the nested container throughout the procedure.

2) Slowly pour or pipette the anticoagulated sample (blood or bone marrow, diluted with saline if needed) along the tube wall into the tube slowly: for a 15 ml separation tube, 4-9 ml of the sample is recommended; for a 50 ml separation tube, 13-30 ml of the sample is recommended.

3) Centrifuge at room temperature with a centrifugal force of 1200 x g for 10 minutes, then turn off the centrifuge. For samples left for more than 24h, longer centrifugation time is recommended.

4) Collect the enriched cells (lymphocytes/PBMCs cells) and pour the supernatant from the separation tube into another clean centrifuge tube, during which the nested container works efficiently to prevent the enriched cells from re-contamination by erythrocytes and granulocytes. Don’t invert the separation tube for more than 2s. (Notes: It is advisable to collect or directly discard the plasma layer of 5-10 mm above the enriched cells before collecting the cells after centrifugation to prevent the enriched cells from re-contamination by platelets.)

5) Wash the enriched cells (lymphocytes/PBMCs cells) with Phosphate Buffer Solution(PBS) then centrifuge them at 250 xg for 10 min.

6) Repeat the washing 2 times as in step 5 and finally re-suspend cells with 5 ml PBS.