Site-Specific Detection of Tyrosine Phosphorylated CD95 Following Protein Separation by Conventional and Phospho-Protein Affinity SDS-PAGE
| Autor: | Sébastien Huault, Krittalak Chakrabandhu, Anne-Odile Hueber |
|---|---|
| Přispěvatelé: | Institut de Biologie Valrose (IBV), Université Nice Sophia Antipolis (... - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Côte d'Azur (UCA) |
| Rok vydání: | 2017 |
| Předmět: |
inorganic chemicals
0301 basic medicine Phos-tag™ medicine.drug_class [SDV]Life Sciences [q-bio] Monoclonal antibody Immunoblot MESH: Phosphoproteins environment and public health 03 medical and health sciences 0302 clinical medicine Protein purification medicine Mobility shift Phosphorylation Tyrosine Receptor Polyacrylamide gel electrophoresis ComputingMilieux_MISCELLANEOUS Death domain MESH: Humans MESH: Immunoblotting MESH: Phosphorylation Chemistry MESH: fas Receptor MESH: Chromatography Affinity Fas receptor MESH: Cell Line 3. Good health enzymes and coenzymes (carbohydrates) 030104 developmental biology Biochemistry 030220 oncology & carcinogenesis MESH: Electrophoretic Mobility Shift Assay CD95 bacteria SDS-PAGE MESH: Electrophoresis Polyacrylamide Gel MESH: Phosphotyrosine |
| Zdroj: | Methods in Molecular Biology ISBN: 9781493967780 CD95 CD95, 1557, Springer New York, pp.173-188, 2017, Methods in Molecular Biology, ⟨10.1007/978-1-4939-6780-3_16⟩ Methods Mol Biol Methods Mol Biol, 1557, pp.173-188, 2017, ⟨10.1007/978-1-4939-6780-3_16⟩ |
| Popis: | International audience; Phosphorylation of two tyrosines in the death domain of CD95 is a critical mechanism in determining the receptor's choices between cell death and survival signals. Recently, site-specific monoclonal antibodies against phosphorylated tyrosines of CD95 have been generated and used to successfully detect each phosphorylated death domain tyrosine of CD95 directly and separately by immunoblotting. Here we provide detailed protocols and useful tips for a successful site-specific detection of phosphorylated death domain tyrosine of CD95 following a protein separation by sizes (conventional SDS-PAGE) and by degrees of phosphorylation (phospho-protein affinity, mobility shift SDS-PAGE). |
| Databáze: | OpenAIRE |
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