Re: Fc Blocking

Rafael Nunez (rafaeln@vetvir.unizh.ch)
Wed, 18 Jun 1997 08:49:11 +0100

Richard
We have been staining human dendritic cell lines and macrophages that
express high Fc receptors, our approach was to use 4% human serum from AB
donors in PBS as blocking and after 30 minutes incubation at 4 degrees
perform staining with either FITC or PE label antibodies in a direct
immunofluorescence assay. It is not perfect but there is a decent
separation from the isotype matched controls.
A tip, if your cells (macrophages) are cultured in medium with red
indicator you might have some additional background that is not due to Fc.
Overnight culture in medium free of red will help with your borderline
markers.
I hope it will work with your system.
Regards and luck
Rafael

>Greetings, everyone:
>
>I am in need of a method to block Fc receptors on human cells. We have
>antibodies (whole molecule) against a ligand on the surface of certain cells
>that we want to use to block binding of those cells to their target - a
>receptor on a human macrophage cell line. However, we need to prevent
>binding to the Fc receptors on the macrophages via the Fc portion of our
>antibodies. We prefer not to have to make F(ab')2 fragments of all of the
>antibodies we will be testing.
>
>I know that anti-CD16/32 monoclonals are available to block Fc receptors in
>the mouse, but as far as I have been able to find out, the anti-human CD16
>and CD32 monoclonals are not efficient as blocking agents.
>
>If you have an efficient method of blocking human Fc receptors, please let
>me know. And, thanks in advance.

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