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: [email protected] [mailto:[email protected]] On Behalf Of WS Sent: Monday, June 20, 2011 5:18 PM To: [email protected] 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 _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods 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. _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods
