I am currently using DIG labelled probes, actually. I have never labelled antibodies before but it may worth a try. The fact is that I would have to design an indirect immunodetection system in order to amplify the signal. The good thing about AP is that it has quite a linear behavior, so the longer your development the higher the signal you will get (under optimal conditions, of course). Besides, I think a homemade version of vector blue substrate would provide a fast and reliable alternative without major changes for many people. Maybe some variation over the naphtol/fast blue system? Thanks ;)
Dear Daniel, If you can make oder get eg DIG labeled probes, it's worth a try: Get some good anti-DIG-AB and label it yourself with activated bright and stable fluorochromes like Alexa or even quantum dots. Preparation is easy, you just need some desalting columns, a photometer is good to have to check the labelling, but not necessary. Another option is an enzyme (AP) conjugate of anti-DIG, together with ELF-phosphate (enzyme linked fluorescence), available from Molecular Probes (now Invitrogen I think), but you'll need a UV light source (390 nm) for excitation (DAPI/H33258 filter set, in that case you may counterstain nuclei with EtBr or PrI). HTH Wo _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods
