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
