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Tyramide Signal Amplification

TSA is a highly sensitive method that allows to detect extremely low-abundance targets in cells and tissues with fluorescence signal 200 times higher than standard imaging methods. This method is easy to use in ICC, IHC, and FISH experimental protocols, using any cell or tissue type. An antibody- or streptavidin-HRP conjugate catalyzes the covalent deposition of fluorescent dye on tyrosine residues of a target protein and antibodies/streptavidin itself. This results in high-density labeling of the target and significantly improves the detection sensitivity.

TSA is a highly sensitive method

TSA is advantageous for fluorescence detection in human tissue, where conventional ICC or FISH often fails to provide adequate signal over autofluorescence background. In applications where increased sensitivity is not required, TSA enables the use of significantly lower antibody amount or probe concentrations for the same level of detection sensitivity thereby reducing issues of non-specific binding or cross-reactivity.

Furthermore, since binding of the tyramide label is covalent, a large number of targets can be detected in the same sample using multiple rounds of sequential TSA, in which the availability of antibodies from different host species is not a limitation. TSA also can be easily integrated with conventional immunostaining.

Human prostate FFPE section labeled with mouse monoclonal anti-pan Cytokeratin (AE1/AE3) antibody and Goat anti-Mouse HRP conjugated secondary antibody following treatment with AF488 tyramide.