Photobleaching Comparison of R-Phycoerythrin-Streptavidin and Streptavidin-Alexa Fluor 568 in a Breast Cancer Cell Line
| Autor: | Jafar Mahmoudian, Seyed Nasser Ostad, Sepideh Babaei, Ali Ahmad Bayat |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Streptavidin Fluorophore Immunology Biotin Breast Neoplasms Heterocyclic Compounds 4 or More Rings 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Immunology and Allergy Humans Sodium dodecyl sulfate Polyacrylamide gel electrophoresis Alexa Fluor Fluorescent Dyes Photobleaching 030102 biochemistry & molecular biology Phycoerythrin Fluorescence chemistry Cell Tracking 030220 oncology & carcinogenesis Biotinylation Biophysics MCF-7 Cells Female |
| Zdroj: | Monoclonal antibodies in immunodiagnosis and immunotherapy. 38(1) |
| ISSN: | 2167-9436 |
| Popis: | Fluorescent dyes are excited by light and emit light at a longer wavelength. Photobleaching is one the most important obstacles in fluorescent image capturing. Photochemical alteration of a fluorescent dye caused by several excitation/emission cycles results in a fluorophore to be unable to emit light. In this study, R-phycoerythrin (R-PE) and Alexa Fluor 568 were separately conjugated to streptavidin. The efficiency of conjugations, R-PE-streptavidin and streptavidin-Alexa Fluor 568, were evaluated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and spectrophotometry, respectively. Herceptin, a humanized therapeutic antibody, was subsequently biotinylated. The reactivity of biotin-labeled Herceptin was examined by enzyme-linked immunosorbent assay. The photobleaching of R-PE-streptavidin and streptavidin-Alexa Fluor 568 were then compared in an immunofluorescent staining on a breast cancer cell line, BT-474. Our data showed that streptavidin-Alexa Fluor 568 was more photostable than R-PE-streptavidin, which provides more time for longer viewing of labeled proteins and image capturing. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|