Combined in situ hybridization and immunohistochemistry in rat brain tissue using digoxigenin-labeled riboprobes

Natasha N. Kumar, Belinda R. Bowman, Ann K. Goodchild

    Research output: Contribution to journalArticlepeer-review

    5 Citations (Scopus)

    Abstract

    This protocol describes a procedure for combining non-radioactive in situ hybridization (ISH) histochemistry with multi-label fluorescence immunohistochemistry (IHC) on rat brain tissue sections. This allows visualization of multiple mRNA and protein targets located within the same or different subcellular compartments. A comprehensive description of RNA probe preparation, validation and storage conditions is described. This is followed by the tissue preparation and tissue processing procedure for combined ISH/IHC on free-floating brain sections. Both the tissue and the RNA probes must be prepared and handle under strict RNase-free conditions up until the point of RNA hybridization.
    Original languageEnglish
    Pages (from-to)31-52
    Number of pages22
    JournalNeuromethods
    Volume70
    DOIs
    Publication statusPublished - 2012

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