I'm calculating amount of sample to apply onto cation exchange chromatography column.
My protein is PDGF (30kDa), which have 24e of net charged. I used Hitrap SP FF column 5ml to purify. In the user guide of GE, they show that the column's capacity is 70mg Ribonuclease A (13.7kDa). I found that the net charged of Ribonuclease A is 29e.
We have a table:
I calculated that: The total charge between two samples is equal. Therefore, x= (70/13.7*29)/24*30 = 179.1 (mg)
I don't know what I do right or wrong? Please help me. Thanks,
* Calculating total charge of protein here: http://nbcr-222.ucsd.../pdb2pqr_2.0.0/
** Getting PDB ID here: http://www.rcsb.org/pdb/home/home.do
Edited by trungnghiatn, 21 April 2015 - 01:16 AM.