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enzyme related question?


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4 replies to this topic

#1 mimiW

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Posted 19 March 2012 - 12:55 PM

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!!!!

#2 mdfenko

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Posted 20 March 2012 - 12:14 PM

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.

Edited by mdfenko, 20 March 2012 - 12:15 PM.

talent does what it can
genius does what it must
i do what i get paid to do

#3 Inmost sun

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Posted 03 April 2012 - 11:51 AM

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

#4 Sid

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Posted 15 May 2012 - 06:31 PM

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

#5 mdfenko

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Posted 16 May 2012 - 07:03 AM

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

in general, fplc column packings are greater in volume than hplc packings. therefore, elution volumes will be at least proportionately greater.
talent does what it can
genius does what it must
i do what i get paid to do




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