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dc.contributor.authorOsman, Tarig Al-Hadien_US
dc.contributor.authorØijordsbakken, Gunnvoren_US
dc.contributor.authorCostea, Daniela Elenaen_US
dc.contributor.authorJohannessen, Anne Christineen_US
dc.description.abstractProtocols for immunohistochemical (IHC) detection of multiple antigens in the same tissue sections have been developed using primary antibodies directly conjugated to different enzymes or fluorochromes, or ones that have been raised in different species, or from different immunoglobulin (Ig) classes or subclasses. For antibodies lacking such dissimilarities, very few proposals have been published with varying degrees of generalizability. In this report we present a successful triple IHC protocol engaging three unconjugated monoclonal primary antibodies raised in the same species and of the same Ig subclass. Compared to other methods, our results showed that denaturation of the preceding reaction complex by microwave heating, combined with additional suppression of enzyme activity, enabled the detection of all three reactions by using the same detection system, with no cross reaction observed. Moreover, expression patterns of each of the three antigens in the triple stained sections, was found to be similar to the pattern observed when single staining was performed. Unlike previous reports, no damage of targeted antigens or tissues did occur following this protocol. Furthermore, the contrast of the colors employed was investigated by computerized color deconvolution, and the three reactions products were successfully separated into three individual images that could be used for further objective quantification.en_US
dc.relation.ispartof<a href="" target="blank">Cancer Stem Cell-Related Markers in Normal and Neoplastic Oral Mucosa. A study on human samples and experimental models</a>eng
dc.rightsAttribution-NonCommercial CC BY-NCeng
dc.subjectTriple immunohistochemistryeng
dc.subjectSame species antibodieseng
dc.titleSuccessful triple immuno-enzymatic method employing primary antibodies from same species and same immunoglobulin subclassen_US
dc.typePeer reviewed
dc.typeJournal article
dc.rights.holderCopyright T.A. Osman et al., 2013. Licensee PAGEPress
dc.source.journalEuropean Journal of Histochemistry

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