title: Detection and isolation of viable mouse IL-17-secreting T cells.
abstract: The MACS Cytokine Secretion Assay technology allows detection of secreted cytokines on the single cell level and sensitive isolation of viable cytokine-secreting cells. In order to label IL-17-secreting cells, a single cell suspension of mouse splenocytes is prepared and stimulated at 37 degrees C with PMA/ionomycin to induce cytokine secretion. To stop secretion cells are then placed on ice and are exposed to the IL-17 Catch Reagent a bi-specific antibody that binds to CD45 on the cell surface of leukocytes and to IL-17 as it is secreted and caught near the cell surface. Secretion is then re-started by increasing the temperature to 37 degrees C and IL-17 is trapped by the Catch Reagent. Secretion is then stopped again, by placing cells on ice. To detect the trapped IL-17, cells are incubated with a second IL-17-specific antibody conjugated to biotin and an Anti-Biotin-PE antibody. Cells can now be directly analyzed by flow cytometry or prepared for isolation and enrichment by subsequent labeling with Anti-PE conjugated MicroBeads.
authors.affiliation: Miltenyi Biotec, GmbH.
authors.forename: Anna
authors.initials: A
authors.lastname: Foerster
authors.forename: Mario
authors.initials: M
authors.lastname: Assenmacher
authors.forename: Michaela
authors.initials: M
authors.lastname: Niemoeller
authors.forename: Elly
authors.initials: E
authors.lastname: Rankin
authors.forename: Mariette
authors.initials: M
authors.lastname: Mohaupt
authors.forename: Anne
authors.initials: A
authors.lastname: Richter
mesh_terms: D000818:Animals; D005434:Flow Cytometry / Q000379:methods; D020381:Interleukin-17 / Q000032:analysis* / Q000378:metabolism*; D015759:Ionomycin / Q000494:pharmacology; D051379:Mice; D012636:Secretory Rate / Q000187:drug effects; D013154:Spleen / Q000166:cytology / Q000187:drug effects; D013194:Staining and Labeling / Q000379:methods; D013601:T-Lymphocytes / Q000166:cytology / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
chemical_list: D020381:Interleukin-17; D015759:Ionomycin; D013755:Tetradecanoylphorbol Acetate