Gel Filtration - which system is most suitable? - (Jan/24/2006 )

Hi,

I have a 18-19kDa protein that appears to be forming higher order structures with itself...possibly a trimer.

I wish to analyse this result further using gel filtration to get better resolution and for purification purposes.

I've been looking through the various gel filtration systems available, namely on the Amersham Biosciences website. There seems to be a number of gel types available such as Sephadex, Sepharose and Sephacryl.

What I would like to know is whether anyone here has used these systems and if one is better than the other?

Thanks in advance,
Ciaran

it's hard to say. i've used two types, but for different purposes (i.e Superdex 200 to deterimine the oligomeric state of a 11kDa protein, Sephacryl 300 for general purification of a 350kDa protein).

i used Amersham's chromatography selection guide to determine what would be best. they also have a Gel Filtration manual that has excellent explanations of the various media types that i found really useful.

hope that helps!

It really depends on how good the separation is.

Generally speaking, Amersham (GE now) provides the best chromatography
system avalible. The column matrixs you mentioned has the follow order
or resolution Sephadex<Sepharose<Sephacryl. Superdex are for high resolution
FPLC in which you need the more expensive AKTA setup to run it.

If you have an ordinary system like the AKTA prime or AKTA FPLC, go for the
Try out the Sephacryl 100 or Sephacryl 200 in a 0.75m or 1m column (self packed).
I think it will give you some good results.

Thanks both of you for replying!!! Appreciate the advice