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 > > . >
