Colloidal gold double labeling of two secretory proteins in bacterial cell
| Autor: | Arati Ghosh, B. K. Ghosh, Guan Tinglu |
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| Rok vydání: | 1983 |
| Předmět: | |
| Zdroj: | Proceedings, annual meeting, Electron Microscopy Society of America. 41:614-615 |
| ISSN: | 2690-1315 0424-8201 |
| DOI: | 10.1017/s0424820100076676 |
| Popis: | Secretory proteins have been localized in bacterial cells by inmunoelectron-microscopy and cytochemical methods. Although ferritin-antibody technique is satisfactory for the localization of single antigenic sites, multiple labelling is not possible by this technique. However, such multiple labelling is possible using colloidal gold antibody because colloidal gold of widely varying sizes can be prepared by simple laboratory manipulation. We prepared colloidal gold of two different sizes and labelled these with anti-penicillinase and anti-alkaline phosphatase antibodies. These labels were used to determine the location of penicillinase and alkaline phosphatase in secreting Bacillus licheniformis 749/C cells.Large colloidal gold particles (150Å) were prepared by 10% sodium citrate treatment of 0.01% HAuCl4; the pH of this colloidal gold suspension was adjusted to 7.6 with 0.2 M K2CO3. Small colloidal particles (50A) were prepared by treating 0.01% HAuCl4, after pH was adjusted to 8.0 with 0.2M K2CO3, withphosphorous saturated ether. The large or small gold particles (1010 in 10 ml) were treated with 100μg of anti-alkaline phosphatase or anti-penicillinase antibodies; after thorough mixing unbound antibodies were removed by centrifugation. |
| Databáze: | OpenAIRE |
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