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Re: GST-Fusion 蛋白提纯求助!!!
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Re: GST-Fusion 蛋白提纯求助!!!# Biology - 生物学
g*s
1
if the protein is purified as GDT-fusion, you must have glutathione in the
prep, then depending on your subsequant application, you may want to dialyze
it using whatever buffer you used to purify it minus glutathione.
the storage question is trickier. if the purified protein is stabel, you can
certainly keep it in -80, but some of them are finicky. temperature shift may
cause aggregation and precipitation after freeze. my feeling is, in protein
manupilation, try to keep as little additives such
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l*b
2
He meaned you don't have to add glycerol at all if you find it does not hurt
the protein property. If you know it is an enzyme, I think you had better add
glycerol, because freezing and thawing cycle can kill the enzyme. For other
proteins, I do not bother to add glycerol either.
And to get rid of gluathione, besides dialysis, there is an easy way. Take a
spin-column with an appropriate cut-off filter, e.g. 30KD, spin, concentrate
the protein and get rid of the solvent in the meanwhile. Then add
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