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IgG
Posted: Thu Mar 10, 2005 10:48 am
by Thomas74
Hi,
has anybody used HPLC to divide labeld IgG from unlabeled?
I´m thankful for every help I can get (e.g. free litreture...).
Sincerly yours
Thomas
Posted: Thu Mar 10, 2005 11:40 am
by HW Mueller
A radioisotope label? Have been trying, but it failed as the medics here don´t think it is necessary to have a gradient cabability. There is that previously mentioned RP-HPLC separation of the different radioiodine insulin which seems to be a starting point for such a trial. You may get lucky with ion exchange, I am to timid to try this, the mixed (up) mode guys might be willing.........
Posted: Thu Mar 10, 2005 11:45 am
by HW Mueller
Well, if you are not so lucky: Maybe the labeling ruined the labeled antibody and it will not be retained on an affinity column.
Posted: Thu Mar 10, 2005 11:54 am
by Thomas74
I´ve a fluorescence labeled IgG (Alexa 532; MW 724)
Posted: Thu Mar 10, 2005 3:04 pm
by HW Mueller
Well that should be a bit easier, but the suggestions hold. Normally the bios are not concerned with the presence of unlabeled stuff, they usually check only whether the labeled antibody is still active. I have done this by incubating the labeled antibody with a tissue slice, then examining the slice via Phosphorimager.