Best Secondary Reagents for Rabbit Polyclonal Abs

Hastings, Richard C richard.hastings at astrazeneca.com
Wed Jun 16 10:40:06 EST 2004


Hi,

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
Associate Scientist
Target Development/TBD
1800 Concord Pike
Wilmington, DE 19803
302-886-2452
richard.hastings at astrazeneca.com




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