Using CPair After One Month

Unused washed CPair can be stored at 4^oC and used for 1 month after washing.

For use after one month, CPair will have to be washed again before use.

1. Tap the CPair tube on the surface to gather all liquid at the bottom of the tube. Do not vortex.
2. Without aspirating all the volume and introducing bubbles, mix the CPair 10x.
3. Do a volume check using an appropriate pipettor to get an estimated volume leftover. Note the value down.
4. Calculate the 90% of the volume and note the value down.
   • Example: For a leftover volume of 100 µL, note down the value 90 µL
5. Place tube onto 1.5 mL tube magnet at room temperature and incubate for 1 min. 
6. Slowly remove 90% of the supernatant, taking care not to disturb the pellet of beads attracted to the magnet.
   • Example: For a leftover volume of 100 µL, remove 90 µL
7. Remove the tube from the magnet and resuspend the pellet by adding 150 μL of cold dPBS, pipetting up and down 5x. 
8. Place tube back on magnet and allow it to pellet for 1 min. 
9. Remove and discard 150 μL of the supernatant, being sure not to disturb the pellet. 
10. Remove tube from magnet and resuspend the pellet by adding the volume removed in Step 6 of cold dPBS into the tube and pipetting up and down 10x. 
    • Example: If 90 µL is removed in step 6, resuspend the bead pellet with 90 µL of dPBS
11. Using the freshly washed CPair beads, briefly mix by pipetting up and down 10x and place the following volumes into each of the pre-coated 0.2 mL Flat-bottom wells
    • For PBMCs/Primary Cells: 20 µL of washed CPair beads
    • For Cell Lines: 15 µL of washed CPair beads + 5 µL of cold dPBS