Unlabeled antibody methods in electron microscopy: a comparison of single and multistep procedures using colloidal gold
| Autor: | Edmund J. Gosselin, J. C. Dennett, G D Sorenson, C C Cate |
|---|---|
| Rok vydání: | 1984 |
| Předmět: |
Calcitonin
Pathology medicine.medical_specialty Histology Serial dilution Immunocytochemistry Antibodies law.invention law medicine Animals Humans Colloids Thyroid Neoplasms Antigens Electron microscopic Antiserum Sheep Chromatography biology Immunoperoxidase Chemistry Goats Middle Aged Microscopy Electron Colloidal gold Immunoglobulin G Immunologic Techniques biology.protein Female Gold Anatomy Electron microscope Antibody |
| Zdroj: | Journal of Histochemistry & Cytochemistry. 32:799-804 |
| ISSN: | 1551-5044 0022-1554 |
| DOI: | 10.1177/32.8.6379035 |
| Popis: | A comparative study of five unlabeled antibody methods was conducted on the electron microscopic level using bridging techniques and colloidal gold. The study was based on the principles of the single-step colloidal gold (GLAD) method (Larsson L: Nature 282:743, 1979) and the multistep single- and double-bridge techniques used in postembedding immunoperoxidase procedures (PAP) (Sternberger LA: Immunocytochemistry, 2nd ed. Wiley, New York, 1979). Using medullary thyroid carcinoma and the same lot of primary antiserum (goat anti-calcitonin) for each procedure, it was shown that adequate localization of calcitonin with the single-step GLAD method was attainable only at dilutions of 1:100 or lower. The single-bridge technique using goat anti-calcitonin, sheep anti-goat immunoglobulin (Ig)G, and goat anti-calcitonin and antigen-coated gold, respectively, worked well at dilutions of up to 1:5000 but not at dilutions of 1:10,000, while single- and double-bridging techniques utilizing goat anti-calcitonin, sheep (Sh) anti-goat IgG, and sheep anti-goat IgG-coated gold produced good localization at a 1:10,000 dilution of primary antiserum. A two-step method using goat anti-calcitonin and sheep anti-goat IgG-coated gold, respectively, appeared to be the most sensitive technique, with adequate antigen localization occurring at a dilution of 1:25,000. While in our hands the two-step method appeared superior in sensitivity to the single-bridge IgG-coated gold technique, each method has its own advantages depending on the individual needs of the researcher. |
| Databáze: | OpenAIRE |
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