Combination of non-isotopic in situ hybridisation with detection of enzyme activity, bromodeoxyuridine incorporation and immunohistochemical markers.
The advent of non-isotopic in situ hybridisation allows the possibility to detect the presence of both mRNAs and other markers in cells. We have established conditions for simultaneous analysis of gene expression and a variety of other immunohistochemical markers in tissue sections. We report the analysis of expression of a family of transcription factors (Sox genes) in combination with detection of: (1) protein antigens (using both monoclonal and polyclonal antibodies); (2) bromodeoxyuridine to mark cells which are proliferating; and (3) acetylcholinesterase activity. The approaches we describe, which demonstrate the compatibility of non-isotopic in situ hybridisation with a range of other treatments, should be generally applicable and open to many variations of probe, antibodies and colour detection systems.
Rex
M
M
Department of Biochemistry, University of Nottingham Medical School, Queen's Medical Centre, UK.
Church
R
R
Tointon
K
K
Scotting
P J
PJ
D000110:Acetylcholinesterase / Q000378:metabolism; D000818:Animals; D015415:Biomarkers / Q000032:analysis; D001973:Bromodeoxyuridine / Q000032:analysis; D002642:Chick Embryo; D004268:DNA-Binding Proteins / Q000378:metabolism; D015870:Gene Expression; D024002:HMGB Proteins; D006609:High Mobility Group Proteins / Q000378:metabolism; D006652:Histological Techniques; D007150:Immunohistochemistry; D017403:In Situ Hybridization / Q000379:methods*; D009363:Neoplasm Proteins / Q000378:metabolism; D009687:Nuclear Proteins / Q000378:metabolism; D055748:SOXB1 Transcription Factors; D055749:SOXB2 Transcription Factors; D055751:SOXC Transcription Factors; D014157:Transcription Factors
D015415:Biomarkers; D004268:DNA-Binding Proteins; D024002:HMGB Proteins; D006609:High Mobility Group Proteins; D009363:Neoplasm Proteins; D009687:Nuclear Proteins; D055748:SOXB1 Transcription Factors; D055749:SOXB2 Transcription Factors; D055751:SOXC Transcription Factors; C490793:Sox11 protein, mouse; C527783:Sox2 protein, mouse; C484490:Sox21 protein, Gallus gallus; C527786:Sox3 protein, mouse; D014157:Transcription Factors; D000110:Acetylcholinesterase; D001973:Bromodeoxyuridine