Best Secondary Reagents for Rabbit Polyclonal Abs

Robin Barclay Robin.Barclay at ed.ac.uk
Thu Jun 17 04:15:49 EST 2004


I have found antibodies from BioSource (www.biosource.com), especially
F(ab')2 goat antibodies - to rabbit or mouse (for MAbs) Ig- to be reliable
for flow cytometry (and blotting) having tried a number of other secondary
antibodies with poor results especially regarding species specificity for
Ig.
Robin Barclay
SNBTS, Edinburgh

----- Original Message ----- 
From: "Hastings, Richard C" <richard.hastings at astrazeneca.com>
To: cyto-inbox
Sent: Wednesday, June 16, 2004 4:40 PM
Subject: Best Secondary Reagents for Rabbit Polyclonal Abs


> 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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