Fluorescence labeling of a cytokine with desthiobiotin-tagged fluorescent puromycin
| Autor: | Takashi Funatsu, Akihiko Sakamoto, Mai Yamagishi, Youichi Aizawa, Takashi Kato, Takafumi Watanabe |
|---|---|
| Rok vydání: | 2008 |
| Předmět: |
Tandem affinity purification
medicine.medical_treatment Cell Biotin Bioengineering Biology Ligand (biochemistry) Applied Microbiology and Biotechnology Fluorescence Cell Line Mice chemistry.chemical_compound Cytokine medicine.anatomical_structure Thrombopoietin chemistry Biochemistry Puromycin Cell culture Fluorescence microscope medicine Animals Fluorescent Dyes Biotechnology |
| Zdroj: | Journal of Bioscience and Bioengineering. 105:238-242 |
| ISSN: | 1389-1723 |
| Popis: | Fluorescence labeling of a cytokine at a specific site is required for observing cytokine-receptor interactions in living cells at the single-molecule level. Here, we demonstrated the C-terminus-specific fluorescence labeling of histidine-tagged thrombopoietin (TPO), a ligand for Mpl, with desthiobiotin-tagged fluorescent puromycin. Fluorescent TPO, purified by tandem affinity purification, stimulated the proliferation of Mpl-expressing cells. Within 10 min of its addition, fluorescent TPO was found to be diffusely distributed on the cell membranes of Mpl-expressing cells, and gradually accumulated to form fluorescent spots. This method is applicable for studying the spatial and temporal distributions of cytokines in individual living cells. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect