Re: Fab2-FITC secondaries
|From:||Gayle Callis <email@example.com>|
Is it adsorbed to mouse? Jackson has a Donkey antiRabbit, adsorbed to
multiple species including mouse, we have used it and their TRITC
conjugates, and used at approx 1:250 to 1:1000 dilution of 0.3 mg/ml. Their
antibodies are very clean, unpricey!
Other possibilities, although an extra step, is an goat xRabbit F(ab')
secondary biotin conjugate adsorbed to mouse (Biosource or Caltag), detect
with SA-Alexa 488 (Molecular probes) superior fluorescence, clean, brighter
and less quenching. SA-FITC is also nice, 488 wins the day. They also sell
Alexa 488 conjugated to polyclonal secondaries, adsorbed to mouse.
Last but more work, buy your favorite F(ab') adsorbed to mouse and use
Alexa 488 conjugation kit, looked easy to do. Molecular Probes kit looked
simple to use.
Good luck --
At 09:44 AM 12/1/00 -0600, you wrote:
>I was quite pleased with my batch of Fab2-FITC (goat
>anti-rabbit) secondary from Boehringer
>Mannheim. Now I'm out. Roche bought up B/M (in the spirit of leaner and
>meaner management style) and Roche no longer sells the antibody. The
>Roche tech rep recommended Chemicon. The best I could get shines real
>green but sticks to the
>mouse tissue I'm working on these days (Head and parts thereof). I use
>plenty of normal serum and 5% BSA so I don't think the blocker is the
>problem. I think the antibody is the problem.
>Does anyone out there have a favorite anti-rabbit IgG to recommend?
>John Carroll Dennis
>Anatomy, Physiology, and Pharmacology
>109 Greene Hall
>Auburn University, AL 36849
Veterinary Molecular Biology
Montana State University
Bozeman MT 59717-3610
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