enzyme related question? - (Mar/19/2012 )

what is the best way to increase my purified enzyme's concentration??? any protocol for concentrating enzymes? also best way to keep them alive n well for the next few month????

thanks!!!!

you can concentrate by ultrafiltration, precipitation and resuspension, evaporation (lyophilization, speedvac, nitrogen stream).

there may be pitfalls to these methods (eg- co-concentration of medium components). you have to see which method will give you stoichiometric recoveries without denaturing the enzyme.

we generally prefer ultrafiltration (with a stabilizing agent when necessary) for final concentration of purified enzyme.

think also of chromatography, I used Q columns to concentrate...

is it true that by using fplc, you'll get more volume instead of hplc?

Sid on Wed May 16 02:31:41 2012 said:

in general, fplc column packings are greater in volume than hplc packings. therefore, elution volumes will be at least proportionately greater.