dallasv at sfu.ca (Dallas Veitch) wrote:
Specifically, we would like the protocol to
>include a gel filtration and an ion exchange chromatogaphy step, and
>to be able to be completed over 3 four hour labs. Does anyone know of
>any protocols which might be useful?
One of the easiest enzymes to purify is the alcohol dehydrogenase from
bakers yeast. The dried yeast is ground up and extracted with phosphate
buffer. An acetone powder is prepared, which is dissolved in phophate
buffer. Further purification step include heat inactivation, ion
exchange, gel filtration and ammonium sulfate crystallization. Assay is
by coupled spectrophotometry. Absolutely studentproof, too.
