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KMID : 1123920040180041154
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Korean Journal of Oriental Physiology and Pathology
2004 Volume.18 No. 4 p.1154 ~ p.1158
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Immunohistochemistry of Paraffin-embedded Tissues by Super-signal Induction Method
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Yun Young-Gab
Lee Jang-Cheon
Jang Seon-Il
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Abstract
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The classical ABC (avidin-biotin peroxidase complex) method for immunohistochemistry in the paraffin-embedded tissues bring into being disadvantage such as low sensitivity of antigen detection and highly background. The biotinyl-tyramide conjugation recently introduced for sensitive immunohistochemistry was applied to light microscopy in paraffin-embedded pancreatic and liver tissues. The protocol consists of an indirect method in which 4-5㎛ tissue sections are reacted successively within a specific primary antibody, followed by a biotinylated secondary antibody, streptavidin-horseradich peroxidase (HRP), and then finally with biotinyl-tyramide. The labeling obtained for insulin and collagen antigen tested in pancreatic and liver tissues, respectively, was found to be highly specific with the labeling for each antigen confined to its particular cellular compartment. In this study, fish (flounder) serum was specially applied to remove nonspecific binding. Background levels and nospecific deposition of the staining were negligible. This results suggest that super-signal induction method by biotinyl-tyramide conjugate can readily applied to antigen detection of the paraffin-embedded tissues.
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KEYWORD
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Biotinyl-tyramide, super-signal, paraffin embedded tissues, insulin, collagen
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