There seem to be many responses to this query, I thought I'ld add some support for biotinylation of the antibody. It's as easy as conjugating to FITC, brighter (because it is a sandwich, and in many cases is brighter even than an immunoglobulin sandwich ... I'ld like to hear if you disagree...), and gets around the problem of "bad" IgG (not-so specific, esp. for B cells). But if you really want to go with a secondary immunoglobulin conjugate, go with an Fab (no cross-linking) or Fab'2 conjugate (test it first). Just tossing this out in hopes it helps, but an ulterior motive is personal edification, so I'm open to "clarifications". Claude.
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