There is a possibility that the reagent ARP http://www.probes.com/servlets/product?region=Select+Region&item=10550 would be useful. This biotin derivative reacts with abasic sites of nucleic acids (nucleic acids that have lost 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=Abstract and abasic sites produced by reactive-oxygen species http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=1567824&dopt=Abstract http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=8347625&dopt=Abstract http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11020331&dopt=Abstract Ionizing radiation also produces abasic sites. Use of a green-fluorescent streptavidin conjugate should work in flow cytometry (or imaging); however, I recommend that you block endogenous biotin first with streptavidin followed by excess biotin. Mitochondria, in particular, have endogenous biotinylated proteins that could interfere with low-level detection. Cells with endogenous damage by ROS may also react as positive so you will need controls. Simon Monard wrote: > Hi > Could you look for chomosome damage by flow cytometry. I don't know how rare chromosome > 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 for abnormal > 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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