Superdex Column - (Nov/08/2006 )
Can any body tell me whats the purpose of superdex column. I guess its used for protein separation according to size of protein if not then what?. and can any body expalin me the no like
superdex 200 16/60 and whats the superdex 200 10/30. and for what purpose column Resource- Q is used.
superdex is a gel filtration packing. it separates by size (larger comes out first, smaller last).
the 16/60 and 10/30 designations refer to column size (1.6 cm x 60 cm, 1 cm x 30 cm).
superdex 200 (as well as 75 and peptide) refers to the exclusion limit of the packing.
resource Q is an exchange column.
It will separate proteins depending on their charges.
First you load the proteins on the column, in a medium containing no salt. then you add more and more salt (for this purpose you use two buffers, buffer A : Tris-HCL and buffer B : Tris-HCl + NaCl 1M, and you switch from 100% buffer A to 100 % buffer so the amount of salt in the column increases, until the proteins will be eluted from the column, and you separate the proteins depending on their charge.
Thats great. Its really a good explanation. thanks for the help