Simon, Roche bases its molecular biology non-radioactive nucleic acid labeling kits on digoxigenin (DIG)-labeled nucleotide analogs, and they have pretty good anti-DIG antibodies to detect these. This system works well in a test tube or on a blot. I don't know whether Roche markets these reagents for cellular DNA incorporation approaches, but I can't think off the top of my head why it wouldn't work just like BrdU. There are anti-BrdU biotin- and PE-conjugates commercially available, and I think the anti-DIG Ab may be available in FITC (?), so you should be able to double-stain. Since you're using it in vivo, the only limitation may cost - I imagine its more expensive than BrdU. (Let me know if you have any luck) Andrew -- Andrew D. Wells, Ph.D. Assistant Professor Department of Pathology and Laboratory Medicine University of Pennsylvania Joseph Stokes, Jr. Research Institute Biesecker Liver Disease Center The Children's Hospital of Philadelphia phone: (215) 590-8710 fax: (215) 590-7384 mailing address: 802 Abramson Research Center 3516 Civic Center Boulevard Philadelphia, PA 19104 On 8/23/02 2:08 PM, "Simon Monard" <smonard@trudeauinstitute.org> wrote: > > Hi > > Has anyone successfully managed to stain cells with bromodeoxy uridine and > iododeoxy > uridine antibodies and been able to distinguish between the two? Seems like it > should > be possible although there is some cross-reactivity between antibodies. If so > which > antibodies did you use and what blocking statagy if any did you use. We want > to challenge > an animal with antigen in the preence of BRDU, let it rest up a while then > re-challenge > with antigen in the presence of IRDU and see if the same cells respond. Can > anyone > think of another way to do this without using radio isotopes? > Thanks for any suggestions. > > Simon Monard > FACS Lab Manager > Trudeau Institute > Saranac Lake > NY12983 > > Ph 518 891 3080 X352
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