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发帖数: 13 | 1 desalting column is much faster, but sometimes proteins bind to it
non-specifically and you may lose some of your sample by that. sometimes,
proteins get denatured because the buffer change is too sudden, not like
dialysis that is slow and "tender". plus, carry over is almost inevitable on
the column, and a 2nd run is sometimes necessary.
dialysis on the other hand, has its disadvantage too. most of all, it takes
way too long (O/N is routine), during which the protein may not be happy.
second, i | n****e
发帖数: 1677 | 2 The salt concentration after the gel filtration should be the same as the
running buffer. I usually use 100mM NaCl to run gel filtration, it depends on
the protein.
I don't know what you gonna do to store the protein you purified. I usually
concentrate the protein down to 10mg/ml, and save it for crystallization.
I save the protein in 5% Glycerol, 10mM DTT, 100mM NaCl and 20mM whatever
buffer. So I usually make the buffer, concentrate the protein down to a
reasonablly small volume, add the buffe |
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