It would be interesting to compare that with the paper of http://www.ncbi.nlm.nih.gov:80/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=9448838&dopt=Abstractas they claim the "Direct detection of 8-oxodeoxyguanosine and 8-oxoguanine byavidin and its analogues" as a measure of oxidative DNA damage. It looks verystraightforward but suggests you should not block with streptavidin but justmeasure it's binding.RegardsGerhard-----Original Message-----From: Richard Haugland [SMTP:richard.haugland@probes.com]Sent: Friday, September 28, 2001 8:20 PMTo: Cytometry Mailing ListSubject: Re: Any thoughts on the detection of Ionizing radiationeffects in ratsThere is a possibility that the reagent ARPhttp://www.probes.com/servlets/product?region=Select+Region&item=10550would be useful.This biotin derivative reacts with abasic sites of nucleic acids (nucleic acidsthat havelost their base but not the backbone structure and thus generate aldehydes.ARP has been used to detect this event in living cells:http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=10639140&dopt=Abstractand abasic sites produced by reactive-oxygen specieshttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=1567824&dopt=Abstracthttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=8347625&dopt=Abstracthttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11020331&dopt=AbstractIonizing radiation also produces abasic sites. Use of a green-fluorescentstreptavidinconjugate should work in flow cytometry (or imaging); however, I recommend thatyou blockendogenous biotin first with streptavidin followed by excess biotin.Mitochondria, inparticular, have endogenous biotinylated proteins that could interfere withlow-leveldetection. Cells with endogenous damage by ROS may also react as positive so youwill needcontrols.Simon Monard wrote:> Hi> Could you look for chomosome damage by flow cytometry. I don't know how rarechromosome> damage would be. Its not very hard to prepare chomsome preps from rats,either con> A stimulated blood or perhaps you could look at granukoma pouch assay cells.Just> a thought. We made some chromosome paints for rat and tried to look forabnormal> chomosomes by microscopy also.> Simon> FLOWers>> I am in need of some help.>> We want to look at the protective effect of drug X on radiation induced> damage in the rat, by Flow Cytometry.> Does anyone have any thoughts??>> I know this is potentially a very broad subject, but I would appreciate any> thoughts on potential avenues of approach> > thank you for your time and help > > Philip Barren
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