And as others have said shedding of dextran is a problem with GE columns
(this was confirmed to me by GE people), but after extensive system
equilibration, we do not see a problem significant enough to ever hurt
our light scattering measurements.
Engin
On 3/8/11 10:38 AM, Engin Özkan wrote:
> On 3/8/11 5:03 AM, Sebastiano Pasqualato wrote:
>> On the other hand, GE Healthcare columns would require a huge amount
>> of material to be loaded.
>>
> What do you mean by a huge amount of material? You would not be using
> a 16/60 column (125 ml column volume) for an analytical experiment.
> How about a Superdex 10/300 (used to be called 10/30) or a 5/150
> column. These have column volumes at 25 ml and 3 ml, respectively,
> have great resolution, and probably already compatible with your
> proteins and buffers.
>
> We used to use HPLC columns, but some proteins would never elute from
> these columns. Then we switched to good old Superdex 200 10/300, and
> it works like a charm every time. We inject <100 ul material at
> concentrations around 1 mg/ml (depending on the molecular weight of
> the protein in question). The only issue is we have to run these
> columns at 0.35 ml/min flow rates (instead of the default 0.5 ml/min),
> since our HPLC has a lot of back pressure for FPLC columns.
>
> Best,
> Engin
>
--
Engin Özkan
Post-doctoral Scholar
Howard Hughes Medical Institute
Dept of Molecular and Cellular Physiology
279 Campus Drive, Beckman Center B173
Stanford School of Medicine
Stanford, CA 94305
ph: (650)-498-7111
|