Hi,
if you really want it SMART was replaced be Äkta Ettan and nowaays
Äkta micro. The nice thing is the easy of use (same software).
However i other think HPLCs might be more flexible and cheaper (e.g.
we have schimadzu - cant complain about anything,
interface is more complex, but you dont really need to care about all
that... you use just a few options - and there are several other
manufacturers..) --- just as a comment on the analytical systems. you
can inject few microliters...
And the ca. 20 ml S-200/S-75 10/300 works fine for small volumes by
the way. at least on our HPLC. why would you need to go smaller??
20-50 ug. in 20 ul should be fine.... did you try? you could try just
chaning the tubing on your current Äktä purifier and injecting
via Hamilto syringe?? Connected to HPLC certainly will work.
Of couse if you want faster runs, thats another thing (i think these
smaller columns are mainly good for fast screening of quality)
HTH,
tommi
On Oct 16, 2011, at 4:14 AM, Artem Evdokimov wrote:
Hi,
You're probably referring to the Superdex 5/150 Tricorn column, with
working volume of 3ml, and not the 15/150?
Those columns work quite nicely for small sample volumes. For
analytical runs 15-25ul injection is pretty nice. The PC columns are
originally designed to be used with the SMART system, which by the
way used to be one of the best analytical products for
macromolecules -- optimized path length, one-volume pumps suitable
for directly running typical protocols, etc. etc. and for its time
the OS was also damn good (OS/2). Sadly, GE did not come up with any
direct replacement for this machine, as far as I can tell. AKTA is
not optimal for analytical runs - tubing is too long, etc.
If you have an old HPLC system moldering in a corner I recommend
either of these columns mounted directly in front of the detector.
In our current setup the entire portion of the HPLC that is
responsible for column selection and heating and so on is bypassed,
so there are literally ~4-5 mm of (the thinnest available PPEK)
tubing in between the injection valve and the column, and in between
the column and the detector. Autosampler is a very helpful feature,
esp. when analyzing fractions output from previous step, and as long
as the column isn't clogged the run is 8-12 minutes (depending on
buffer composition). Now, Agilent software for HPLC is absolutely
horrible for this kind of work but it suffices.
Artem
P.S. for lower protein quantities don't forget to record the A210,
in addition to A280 and A260.
On Sat, Oct 15, 2011 at 5:04 PM, Alexandra Deaconescu <deac...@brandeis.edu
> wrote:
Hi everyone:
I was hoping I could get your opinion on the performance of the
small approx. 3ml gel filtration columns from GE Healthcare. We
currently have an Akta fplc and we would like to do small runs using
small volumes of sample. As far as I know we have two options:
1. use the Superdex 15/150 columns
these appear to have a slightly lower resolution (the no. of
theoretical plates is about 25000 m-1)
these apparently can be directly hooked up to our Akta fplc
2. use Superdex PC 3.2
these have somewhat higher resolution (the no of theoretical plates
is 30000 m-1)
these can only be hooked up using a special "Precision Holder"
equipped with titanium end fittings (which I have heard clog easily?)
What is your experience with these columns?
Which option of the two do you recommend (cost is not a huge issue,
but resolution and overall performance is)?
Have you seen significant band broadening when using these small
columns with a regular fplc (rather than Akta's microFPLC)?
I would greatly appreciate your comments! Many thanks...
Bests,
Alex
.
Tommi Kajander, Ph.D.
Structural Biology and Biophysics
Institute of Biotechnology
University of Helsinki
Viikinkaari 1
(P.O. Box 65)
00014 Helsinki
Finland
p. +358-9-19158903
tommi.kajan...@helsinki.fi