APOLLO QUICK START GUIDE
If in doubt please consult the full 20 ml Apollo or 7 ml Apollo User Guides, which are only summarised here for new users to easily gain familiarity.
HOW TO USE APOLLO CONCENTRATORS
Apollo concentrators have a very fine filter with surface pores which reflect the protein, viral gene particle, or macromolecule of interest that you want to concentrate in the top (this solution is called the retentate). Molecules much smaller than the pore size can pass freely through the filter and end up in the collection tube (this solution is the filtrate).
Protocol:
1 Select a preassembled device consisting of collection tube and Apollo filter that has a molecular weight cut-off (MWCO) equal to or smaller than the native MW of your protein of interest.
2 Check the full Apollo User Guide* to confirm that the device is compatible with any harsh chemicals you may be using e.g. certain organic solvents, concentrated NaOH.
3 Make sure that it will fit your centrifuge – you will probably need a carrier tube or insert to fit the device into the rotor or swing bucket/head before you start centrifugation. Fixed angle centrifuges or swing bucket/head centrifuges can be used, but best results are obtained with a swing bucket/head. Check that the there is sufficient clearance above the Apollo concentrator for the centrifuge to spin normally.
4 If you need to remove glycerin (a stabilizer present in the Apollo membrane) then add some clean water and spin (using a balance tube) at the recommended relative centrifugal force (rcf) to pass some water through the membrane*. Shake the water out of the device and reassemble.
5 Add sample and cap tube tightly.
6 Place assembly in rotor with balance tube and spin, being careful to observe centrifugal force limits for both the Apollo device* and the rotor you are using. Spin times vary depending on the solution you are using and the desired degree of concentration. You can stop the spin after e.g. 5-10 min and estimate how long you will need to spin for. Some users check how fast concentration is proceeding at various points in the run.
7 Harvest your concentrated retentate with a 200 uL or smaller pipette tip to avoid damage to the membrane near the tip of the concentrator. Viewing from above, depress to the first stop, slide tip gently down the groove formed by the vertical membrane seam, and aspirate retentate from the concentrator. You can improve recovery of any protein film left in the device by rinsing just the the tip of the membrane with 5-50 ul buffer, optionally allowing it to stand for a few min before removing the rinse buffer.
TIPS:
• Avoid scraping the membrane with pipette tip
• Avoid excessive spin speeds – especially with high MWCOs
• For best recovery remove your concentrated solution from device in <10 min
* See full protocol for details.