Gel Filtration Chromatography Protocol

Biochemistry, Molecular Biology, and Cell Biology Protocols >> Gel Filtration Chomatography Protocol

Running a size exclusion column

Applying the sample
Pour beads down a glass rod (placed in the middle of the tube)
Let the beads settle, and then connect the top reservoir of buffer to it
Wash buffer two more times (they recommend seven volumes of the beads for washing initially and washing between samples)
Measure the bed volume (i.e. the height of the beads)
Let buffer run to the top of the column
Stop the column
Place rubber tubing around Pasteur pipet
Slowly apply sample down side of the column
Let sample run to the top of the column
Stop the column
Add a bit more buffer
Let column drain down
Then, apply enough buffer and start connecting the reservoir again
Collect fractions