Hello,
Has anyone used an alexa labeled (or any other fluorophore) anti-DIG or anti-FAM antibodies for Locked Nucleic Acids (LNAs) for ISH from Exiqon? Exiqon does not provide LNAs labeled for fluorescence (as far as I know) and I would like to use some alexa but I don´t know if this may work with miRCURY LNAs.
Thank you, hope you can help me
Hi Rafael, I have checked the manual of this ISH kit. There are some detection possibilities mentioned. I would recommand the detection with Sheep FAB Anti-Dig.POD (Roche cat# 11207733910), dilution between 1:100 and 1:400. After 90 min incubation do the biotinylated Tyramide step and then you should detect this bindings with StreptAvidin-Fluorophore (CY2,3 or 5 or Alexa dyes). The tyramide-technique is an additional enhancement step. We use this ISH protocl on 25 µm thick native cryo sections.
Other possibility: take an Anti-DIG H&L antibody and detect this binding either with a fluorophore labelled secondary antibody or if this is not sensitive enough use a biotinylated secondary ab and then again a StreptAvidin-Fluorophore. Good luck!
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