need protocol on purfication of allinase from raw garlic - (Jan/04/2011 )
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.. =>
-ikwana-
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).
-mdfenko-
mdfenko on Thu Jan 6 21:26:47 2011 said:
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.sciencedirect.com/science?_ob=ArticleURL&_udi=B6WB5-460Y3X5-5&_user=2512477&_coverDate=06%2F15%2F2002&_rdoc=1&_fmt=high&_orig=search&_origin=search&_sort=d&_docanchor=&view=c&_searchStrId=1599181727&_rerunOrigin=google&_acct=C000027478&_version=1&_urlVersion=0&_userid=2512477&md5=58a429e7ae8fdcc50048712817c5d4b8&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 =>
-ikwana-