Matt Gethers/CRI, Thailand/Labwork/Concentration of Protein Sample
Concentration of Protein Sample through Millipore Centrifugal Filters
Using the Amicon Ultra-15, load as much as 15 ml of sample and spin at 4,000 g for 20 minutes at 4oC. You can spin for additional time to further concentrate the sample left between the filters. When at the desired volume, pipette out the sample into an eppendorf tube.
Ran the q-sepharose purified HmgR on the column in two rounds of 20 minutes at 4,000g at 4oC. I then spun down the volume remaining in the chamber between the filters for 15 minutes at 4,000g and 4 oC. I was left with ~400 μl of sample. I saved all the flow through in case the column failed to bind the HmgR.