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KMID : 0378019700130050054
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New Medical Journal
1970 Volume.13 No. 5 p.54 ~ p.58
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Standardization of Enzyme-labeled Antibody Method
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羅鳳鎭/Rha, Bong Jin
李洸鎬/丁炳相/白相豪/Lee, Kwang Ho/Chung, Byung Sang/Baik, Sang Ho
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Abstract
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Under various status of fixation, washing, substrates and incubation, several conditions for an application of enzyme-labeled antibody method to localize LH in the rat pituitary gland were obtained and discussed.
Bouin´s solution could preserve endogenous LH most effectively after a longer duration of fixation and washing than that requires ordinarilly.
Intense reactions of the specific LH cells were observed with 3, 3´-diaminobenzidine as a substrate, while a-naphthol and 4-Cl-l-naphthol showed lower intensity in either of specific cells or nonspecific backgrounds.
The most and the only adequate concentration to reveal the intense staining was found at 0.001% of hydrogen peroxide in substrate solution.
Residual of fixative chemicals in the tissue, higher concentration of hydrogen peroxide and longer incubation in substrate solution may act as a factor or factors to develop nonspecific staining of background tissue.
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