Toyopearl HW-55 is for rather large proteins, HW-45 might be more appropriate (but you can not know until you try...).

Packing SEC columns is an art (to avoid preferential pathways) but it is indeed perfectly possible to carry out SEC using glass columns and gravity. That's how it was done until the 90's. I have actually done it using just a (very long) glass cylinder with stopper (with a steel tubing inside), a bit of nylon cloth, a few glass beads and glass wool for the lower end of the column, and just a stopper with a hole and steel tubing on top. There was at that time a VERY GOOD booklet from Pharmacia, maybe somebody still has a copy somewhere.

You might use our PSS PROTEEMA columns. The problem is that usually do not know the molar mass of your Protein an thus the required pore size of the column in advance (see comment of Catherine above). In addition, depending on you protein (mainly PI and polarity), the eluent migh vary.

Therefore, we offer a column selection service at low costs. This Service includes running the sample on our column and suggesting  a suitable eluent with column (combination) such that you sample neither elutes too close to the exclusion limit nor at the separation limit of the column combination.  

Provided we can identify suitable conditions and you buy the column, the costs for the column selection Service will be deduced from the price of the column.

This is not a complete methode development, but provided a good staring point 

For more Information, you may contact my collegue Jochen Leinweber at [email protected].

Best regards

Wolfgang

Thanks a lot Catherine and Wolfgang,

Catherine - finally I decided to use Sephacryl S-300 and glass column (50 ml, 12 mm) with stopper. Seems will be hard to found Pharmacia book you've recommended - I put glass wool to the lower end of the tube - could you tell why (or do)  we need glass beads you've mentioned and if we need - what kind of beads it should be? 

BR

German