remove BSA from IgG prep
Posted 10 November 2004 - 04:43 PM
i am trying to purify a rat monoclonal antibody from tissue culture sup (1%FBS). after ammonium sulfate precipitation (50% fraction) i resuspended the pellet in PBS and dialysed against PBS to get rid of the salt. on SDS PAGE it looks like it has so much of albumin in it.
i tried affi-Gel blue gel from bio rad but it is not working well for some reason. i did equilibrate my sample in phosphate buffer. with proteinG, the yield was very low when i tried directly with TC sup. probably not all of them got eluted even after repeated elution. gel filtration (sephadex G100) does not separate them far enough. i have not tried ion exchage.
thanks in advance.
Posted 11 November 2004 - 07:53 AM
Try protein G or A Sepharose from Amersham-Pharmacia. It should bind just your Ig, but not other proteins.
Posted 17 December 2004 - 02:01 AM
best of luck