Using the Stirred Cell Concentrator: Difference between revisions
From Powers Wiki
(Created page with "Using the Stirred Cell Concentrator # Obtain a 44 mm membrane for the concentrator. Handle the membrane by touching only its edges. # Place the membrane, shiny side down, in...") |
(Added to Category) |
||
Line 22: | Line 22: | ||
# Turn the pressure release valve to the horizontal position to relieve pressure. | # Turn the pressure release valve to the horizontal position to relieve pressure. | ||
# Disassemble and clean. | # Disassemble and clean. | ||
[[category:Protocols|Protocols]] | |||
[[category:Protein_Preparation]] | |||
[[category:General_Maintenance]] |
Revision as of 03:14, 11 September 2021
Using the Stirred Cell Concentrator
- Obtain a 44 mm membrane for the concentrator. Handle the membrane by touching only its edges.
- Place the membrane, shiny side down, in a beaker of nanopure water (100 mL).
- Change the water in the beaker three times over the next hour. This ensures that the storage buffer is completely removed from the membrane before use.
- Collect all of the parts needed to assemble the concentrator. Consult the manual if necessary.
- Lubricate the two red O-rings adequately. You should be able to slide your gloved fingers easily around them.
- When the membrane is clean, place it shiny side up on the grooved bottom piece of the concentrator.
- Place a red O-ring on the top of the membrane. Be sure to not get any lubricant all over the shiny surface.
- Place the cell body over the grooved piece and membrane and screw on the bottom plate. Ensure that the cell is sealed but do not over tighten.
- Place the stir bar insert into the concentrator cell.
- Insert the sample that is to be concentrated into the cell. Do not exceed the 50 mL mark.
- Slide the top of the concentrator onto the cell. It should go on easily. Forcing the lid on will dislodge the O-ring. Apply more lubricant to the O-ring if necessary.
- Turn the pressure release valve (black lever) on the cap to the horizontal position.
- Screw the gas inlet onto the cap. Do not over tighten.
- Place the cell into the retaining stand and turn the pressure release valve to the vertical position. Place the cell and stand on a stir plate.
- Turn on the air pressure slowly.
- Set the regulator to 50-55 psi. Do not let the pressure exceed 75 psi.
- Once the cell is pressurized, start stirring. The vortex should go down 1/3 of the total volume of the liquid.
- Concentrate to a volume greater than 2.5 mL.
- Turn off the air pressure.
- Turn the pressure release valve to the horizontal position to relieve pressure.
- Disassemble and clean.