volume of sample to load on column??

[Farah Saleem]

Dear all,

I am working on the separation of bacterial beta galactosidase enzyme from the mixture of protein sample. Doing conventional gel filteration chromatography for this purpose. Resin is Sephadex G-100.
Kindly suggest me how much volume of sample should i load on column?? My sample contains 1.2 mg/ml of protein.
Column height is 80cm and width is 2.2 mm.

Kindly reply as soon as possible.

[Kristof]

Less than 5% of column volume, around 1-2% should be OK.

[ScottHorn]

Double check those column dimensions. Are you sure you didn't mean 80cm by 22mm? Otherwise that sounds like a very impractical column for your purpose, which I'm assuming is prep scale. Also, that would be an incredibly fragile column.

[Farah Saleem]

thanks for your reply