Immunochemistry question (probably dumb)
(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.
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)
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
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.
Methods mailing list
Methods from net.bio.net
This email message may contain legally privileged and/or confidential information. If you are not the intended recipient(s), or the employee or agent responsible for the delivery of this message to the intended recipient(s), you are hereby notified that any disclosure, copying, distribution, or use of this email message is prohibited. If you have received this message in error, please notify the sender immediately by e-mail and delete this email message from your computer. Thank you.
More information about the Methods