Best Secondary Reagents for Rabbit Polyclonal Abs
g.nebe-von-caron at unipath.com
Thu Jun 17 10:05:18 EST 2004
At last not a virus.
I actually also use the Jackson donkey ant rabbit but in PE for surface
fluorescence with great success. Have previously used the sigma rabbit
anti-FITC monoclonal trying to follow some published protocol but that
was very poor in comparison. Have not tried the donkey FITC reagent.
From: Hastings, Richard C [mailto:richard.hastings at astrazeneca.com]
Sent: 16 June 2004 16:40
Subject: Best Secondary Reagents for Rabbit Polyclonal Abs
Lately, I have been facsing many rabbit polyclonal antibodies. I have
been using Molecular Probes goat anti-rabbit IgG Alexa Fluor 488 and
Jackson Immunoresearch donkey F(ab')2 anti-rabbit IgG (H+L) FITC or APC
as secondary reagents. I use fluorochromes that don't overlap with
propidium iodide because I like using PI as a marker of cell death
(yeah, I know PI is more a marker of membrane permeability, so humor me
a bit here and I do know that APC and PI overlap in their emission
wavelengths). Problems I have run into are these secondary reagents are
not bright at all, the rabbit IgG preps tend to have a lot of
non-specific binding and purified rabbit IgG is a lousy control.
I have also tried the Molecular Probes rabbit Zenon kit and the results
were less than satisfactory. Anyone have any suggestions?
Richard C. Hastings
1800 Concord Pike
Wilmington, DE 19803
richard.hastings at astrazeneca.com
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