From owner-drosophila@net.bio.net Wed Dec 21 22:00:00 1994 Path: biosci!agate!howland.reston.ans.net!vixen.cso.uiuc.edu!news.uoregon.edu!bappel.uoregon.edu!user From: appel@uoneuro.uoregon.edu (Bruce Appel) Newsgroups: bionet.drosophila Subject: Re: Fluorescent In situ Date: 22 Dec 1994 20:41:24 GMT Organization: University of Oregon Lines: 33 Distribution: world Message-ID: References: <94Dec22.095229est.18900@gpu.utcc.utoronto.ca> NNTP-Posting-Host: bappel.uoregon.edu In article <94Dec22.095229est.18900@gpu.utcc.utoronto.ca>, ilivne@UTCC.UTORONTO.CA ("I. Livne") wrote: > Dear netters > Has any one tried the Molecular Probes ELF substrate for detection of Digoxigenin probes > in whole mount in situ hybridization? On paper it looks as promising alternative to the > NBT substrate and generates clean Fluorescet signal. Is it for real? > > Livne Bar > I.Livne@utoronto.ca We've tried out the ELF substrate to detect several different digoxygenin-labeled probes hybridized to whole-mount zebrafish embryos. We find that it is less sensitive than NBT/BCIP and even slightly less sensitive than Fast Red. In addition, the reaction product forms a crytalline precipitate, which is not evenly distributed throughout the cell, thus reducing cellular resolution. Finally, the substrate comes out of solution after about 2 hr and sticks to the tissue, causing additional problems. Despite the problems, I can see it having some utility. For example, if you want to mark a domain (as opposed to individual cells) with a fluorescent signal, this may do it for you. Hope this helps. Bruce Appel -- Bruce Appel University of Oregon appel@uoneuro.uoregon.edu .