Immunochemistry question (probably dumb)

Jayakumar, R via methods%40net.bio.net (by R.Jayakumar from roswellpark.org)
Tue Jun 21 08:49:36 EST 2011


Secondary antibodies may have also been adsorbed against each other to ensure specificity before being recommended for double staining.  As Wo suggested, (Fab)2 fragments missing their Fc fragments but directed against FC of primary may also be used to prevent non-specificity.  OR IT MAY JUST BE A TYPO error.  
  
Jay


-----Original Message-----
From: methods-bounces from oat.bio.indiana.edu [mailto:methods-bounces from oat.bio.indiana.edu] On Behalf Of WS
Sent: Monday, June 20, 2011 5:18 PM
To: methods from magpie.bio.indiana.edu
Subject: Re: Immunochemistry question (probably dumb)

Dear Peter,

this question is not dumb. If complete antibodies were used, one
should have amplified the other (and vice versa). That's what people
usually do to amplify signals!

Apparent differences in the stained structures (I assume some sort of
detection by photography, video or confocal scanning microscopy),
simply may arise from different sensitivities in the optical channels
used.

One possibility I can imagine where such an experiment gives reliable
results is, when the secondaries are directed against the other
species' Fc, but missing their own Fc (means they are incomplete
antibodies), which might be quite unusual/uncommon.

HTH

Wo
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