what is the diameter and length of the column?
1ml/min sounds a bit quick for GPC
John Fox
> So I have been having problems running my gel filtration column. It is a
> High Resolution column put out by Amersham Pharmacia. Before loading my
> protein I equilibrate the column with 3 column volumes of buffer. This
> buffer among other things contains 150mM salt per the instructions to
> eliminate nonspecific interactions of the protein with the column. Then I
> load my sample which has been concentrated down to under 1 mL and run the
> column at 1 ml/min. I collect about 2 mL fractions. When I run an SDS-PAGE
> gel of these fractions I see proteins of all molecular weights in each
> fraction, reflecting poor separation. It's as if I never put the protein
> sample through the column. I feel like it must be something I am doing
> wrong and not the column because at least one other member of my lab has
> used this column and achieved nice separation. Any thoughts on what the
> problem might be?
>> Erin Warren
>> <http://www.biowww.net/forum/read.php?f=6&i=999&t=999>
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