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need protocol on purfication of allinase from raw garlic


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#1 ikwana

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Posted 04 January 2011 - 09:51 PM

i need to isolate enzyme allinase from raw garlic. i need to use column chromatography.. what reagent i have to mix with the raw garlic ( for crushing, blend, homogenization)before pouring into column? what kind of gel that i have to use in the column? how am i gonna isolate only the allinase?

thank u.. =>

#2 mdfenko

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Posted 06 January 2011 - 01:26 PM

that depends on what your first column will be.

if it is gel filtration then pbs or tbs should be okay (you may want to add 0.5-1mM dtt to keep sulfhydryls reduced).

if it is ion exchange then you may want to use 20-50mM tris or 10-20mM phosphate at an appropriate pH (with dtt).

Edited by mdfenko, 06 January 2011 - 01:27 PM.

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#3 ikwana

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Posted 07 January 2011 - 12:48 AM

that depends on what your first column will be.

if it is gel filtration then pbs or tbs should be okay (you may want to add 0.5-1mM dtt to keep sulfhydryls reduced).

if it is ion exchange then you may want to use 20-50mM tris or 10-20mM phosphate at an appropriate pH (with dtt).


hey,thank u 4 ur concern..

owh... i so the gel filtration.. but i've the protocol to purify the enzyme (alliinase)

this the link of the journal
http://www.sciencedi...b8&searchtype=a


to blend it,
i have all the material, except
-pefabloc,PLP,PEG-800, methyl-alpha-D-mannoside, HiLoad 26/60 superdex 200 prep grade,alpha-globulin, racemic alliin,lactate dehydrogenase, NADH

do u have any idea, any other chemical that can replace these?

thank u =>




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