Re: Double labeling BrdU/Ki-67 in rabbit

From:"John C. Dennis"

Hey Pat

In re your #2:

I can't say that a sheep antibody  would make a difference although the
particular antibody you have may not be the best.  It's probably just

Your background may depend on whether or not you have the right dilution
of the primary.  Polyclonals are particularly sensitive to dilution.  Have
you run a dilution series all the way out to invisibility?  For example,
I'm using some polyclonals at 1:16,000.  I never expected that sort of
skookum  response and it took me three runs to go that far out in to
Dilution World.

You didn't mention how you're trying to visualize the BrdU so we have to
assume you ran the right blocking steps?

I grew up in Big d.  Do the egrets still roost in the trees by the med

John Carroll Dennis
Anatomy, Physiology, and Pharmacology
109 Greene Hall
Auburn University, AL  36849

On Tue, 22 Jan 2002, Patrick Ladage wrote:

> Hi,
> Please advise: I would like to double label BrdU/Ki-67 in the rabbit cornea, preferably en bloc and otherwise in cryostat sections.
>  (1)  Is there a Ki-67 antibody that works in the rabbit and that is not a mouse monoclonal?
> (2)  What about BrdU? I tried sheep polyclonal but it gives too much nonspecific background staining.
> (3)  Do you have some references on double labeling BrdU/Ki-67 in the rabbit.
>  Thanks for your help,
>  Patrick
> *****************************************************
> Patrick M Ladage B.Optom F.A.A.O.
> Jr. Postdoctoral researcher
> Department of Ophthalmology
> UT Southwestern Medical Center, Dallas, TX
> 5323 Harry Hines blvd, Dallas, TX 75390-9057
> (P) 214- 648- 9544
> (F) 214- 648- 8447
> Ph.D. candidate
> College of Optometry
> University of Houston, Houston, Texas
> *****************************************************

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