Re: IgM antibodies and ugly histos
Scott I. Simon (ssimon@bcm.tmc.edu)
Wed, 28 Feb 1996 10:40:52 -0600 (CST)
Dear Steve;
In looking at adhesive interactions using IgGs and IgMs there is a big
difference in the ability of the two to induce aggregates. IgM has
multiple binding sites and although each site may be of low affinity or
even down to nonspecific levels of affinity they cause big time
aggregation of cells. You want to observe binding to singlets alone and
there are at a minimum two alternatives. Gate on singlets (assuming there
are enough of them) using another channel such as SSC vs FSC or an
irrelevant binding mAb or dye (IgG!). The second is to crack the IgM into
Fabs and hope it still has a high enough affinity for detection. We call
it the IgM blues.
Good luck
Scott Simon.