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

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

doi:10.1007/978-1-61779-897-9_2

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

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

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6 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 language	English
Pages (from-to)	31-52
Number of pages	22
Journal	Neuromethods
Volume	70
DOIs	https://doi.org/10.1007/978-1-61779-897-9_2
Publication status	Published - 2012

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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.",

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AU - Goodchild, Ann K.

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N2 - 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.

AB - 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.

UR - https://www.scopus.com/pages/publications/84982873618

U2 - 10.1007/978-1-61779-897-9_2

DO - 10.1007/978-1-61779-897-9_2

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SN - 0893-2336

VL - 70

SP - 31

EP - 52

JO - Neuromethods

JF - Neuromethods

ER -

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

Abstract

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