Tyramine Amplification Technique in Routine Immunohistochemistry

Abstract

SUMMARYSignal amplification in immunohistochemistry via binding of biotinylated tyramine to proteins near the site of peroxidase-labeled antibodies is a promising new technique, but studies investigating a wide range of markers are lacking. The tyramine amplification technique (TAT) was investigated on 85 antibodies using a simple and fast protocol, and TAT results were compared to those obtained with conventional immunohistochemistry. Using TAT, most of the markers could be 5-to 50-fold further diluted and still showed identical staining results compared with standard stainings (maximal 500-fold). However, the variable reactivity of the different markers with TAT underlines the need for individual testing of every antibody to determine the optimal dilution. Some antibodies against cell adhesion molecules could be demonstrated for the first time in archival, formalin-fixed tissue sections. TAT, if carefully evaluated, offers a revolutionary improvement for modern immunostaining, either to increase sensitivity or primary antibody dilutions (cost reduction). From a methodological point of view, immunohistochemistry has not reached its limits by far and TAT is an important progressive step in this developmental process.(

Kaposi's Sarcoma-Associated Herpesvirus Promotes Angiogenesis by Inducing Angiopoietin-2 Expression via AP-1 and Ets1

“…After microwave pretreatment (pH 7.2) for epitope retrieval, 3-or 4-m serial sections from paraffin-embedded, formalin-fixed KS lesions were stained with an affinity-purified goat polyclonal antibody to Ang-2 (R&D Systems, Abingdon, United Kingdom) at 1:100 dilution or to an isotype control antibody (R&D Systems). The primary antibodies were then revealed with an anti-goat-horseradish peroxidase conjugate (R&D Systems), and the signal intensity was increased by the tyramine amplification technique, as described previously (51,71). To detect LANA, a rat monoclonal antibody to LANA (Novocastra Laboratories, Newcastle Upon Tyne, United Kingdom) was used at 1:750 dilution, and the signals were revealed with an anti-rat secondary antibody-horseradish peroxidase conjugate (R&D Systems).…”

Alteration of subcellular and cellular expression patterns of cyclin B1 in renal cell carcinoma is significantly related to clinical progression and survival of patients

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Sensitive detection of GFP utilizing tyramide signal amplification to overcome gene silencing

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