Packing of cartridges with purification resin

This protocol describes how to pack a cartridge / column with protein purification resin. It applies to all agarose resins that are avalible in our store.
The empty cartridges / columns that this procotol uses as a reference are listed here.
To composition of the agarose storage buffer can be found here.

Protocol:

Packing of cartridges /columns with protein purification resin
Recommendation: Since this is a process that requires some handwork, we prepared a video for you that visualizes the packing process. Have a look at it, before packing your first cartridge.
 
Required equipment:
  • Empty tube
  • Sealing sleeve
  • Bottom stop plug
  • Top and bottom frit
  • Top stop plug
  • Sealing plug
  • Syringe
  • Cartridge holder
  1. Push the smaller frit into the empty tube.
  2. Shake the resin bottle to mix beads with storage buffer. This has to be done because otherwise the beads settle on the bottle bottom.
  3. Fill a syringe with resin storage buffer and ensure that no air is left in the syringe. It is essential to avoid any air bubbles in the purification cartridge / tube from now on. This applies to all steps up to step 9.
  4. Fill approx. one quarter of the empty tube with agarose storage buffer. Again, ensure that no air bubbles are in the buffer.
  5. Add 500 µl mixed purification resin to the column. Make sure that the resin slurry is still properly mixed and the beads did not settle.
  6. Remove most of the storage buffer through the use of the syringe. But do not leave the resin beads dry! Otherwise air bubbles might form in between the small agarose beads.
  7. Add another 500 µl of purification resin. Again ensure that it is still properly mixed.
  8. Top up the tube with resin storage buffer. Up until it starts to flow over. No possible space for air bubbles should remain.
  9. Add top frit. It is normal that some of the just added storage buffer spills over here. This is actually a good sign because it means no air was left in the tube.
  10. Add the sealing plug and place the tube in the cartridge holder.
  11. Apply pressure on the cartridge to heal the resin airtight between the two frits. Again it can occur that some form of liquid spills over when the pressure is added.
  12. Screw on the bottom stop plug and add the sealing sleeve.
  13. Complete column by adding the top stop plug.

Agarose Storage Buffer

 
ComponentConcentration
Sodium Acetate 20 mM
Ethanol 20%
Instructions: Add water to your total volume.
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