Protocol Online logo
Top : Forum Archives: : Immunology and Histology

Mouse IgG purification - Whether to use Protein A or Protein G column (Aug/12/2006 )

Hi all,
I have few monoclonal antibody secreting clones against one of the mycobacterial antigens.
I would like to purify few of them from the culture supernatants to do an assay with them.
I was confused whether to use a protein A or a Protein G column for the purification of mouse IgG from the tissue culture supernatants having 10% FBS. We have protein G HiTrap column but only protein A beads.
Any suggestions would be grateful
Thanking you in advance
Leelaram

-leelaram-

You can check here the affinities of protein A and protein G for different isotypes of immunoglobulin.
I would choose protein G

http://www.merckbiosciences.co.uk/html/cbc...einG.htm#table1

-Missele-

I would use protein A instead of Protein G, as FCS will bind to Protein G.

I hope this may help.

-Minnie Mouse-