IgG

[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

[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.........

[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.

[Thomas74]

I´ve a fluorescence labeled IgG (Alexa 532; MW 724)

[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.