Re: BrdUTP-conjugates

From: Mark Shlomchik (mark.shlomchik@yale.edu)
Date: Tue May 16 2000 - 16:18:06 EST


At 9:39 AM -0700 5/15/00, Barsky, Lora wrote:
>Hi Everyone,
>
>I'm getting a protocol started here and will need to use BrdUTP for a
>proliferation assay.  Will also be looking at PKH26 and GFP, so both FITC
>and PE conjugates to BrdU cannot be used.  Has anyone used the Cascade Blue
>(UV excite) or are any RED colors (488 excite) available as a conjugate or
>an indirect stain with BrdU?
>
>My systems are a Vantage with 488 argon, 350-361 UV, and a 632 HeNe.  And a
>4 Color Calibur (+HeNe) and a 3 color Scan.
>
>Would staining for BrdU always work best, like other cytoplasmic stains,
>when FITC is the color?
>
>Thanks,  Lora Barsky

Check out the products made by Phoenix Flow. They work very well, are
much cheaper than the BD, and also titer out further in our hands.
Their Ab (PRB1) is the only other one I know of besides the BD Ab
that works well with the DNAse tx protocol.  They have the Ab
conjugated in several colors. We have used the FITC and in particular
the biotin, which we developed with APC. The latter gave us the best
separation on a FACScalibur but also works on a FACS Vantage with
HeNe laser.

If anyone knows of other mAbs, particularly in the public domain,
that are compatible with the DNAse method of detecting BrdU, please
let everyone know.

Mark Shlomchik, MD, PhD
Associate Professor of Laboratory Medicine
and Immunobiology
Yale University School of Medicine

203-688-2089
203-688-2748 (fax)
mark.shlomchik@yale.edu



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