Multicolor flow cytometric analysis of IL-17 Receptor B on innate lymphoid cells in mouse bone marrow. BALB/c mouse bone marrow cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) (Cat. No. 553141 or 553142). The cells were then stained with BD Horizon™ V450 Mouse Lineage Antibody Cocktail (Cat. No. 561301), FITC Rat Anti-Mouse CD25 (Cat. No. 553071), PE Rat Anti-Mouse CD127 (Cat. No. 552543), and either Alexa Fluor® 647 Rat IgG1, κ Isotype Control (Cat. No. 557731, left panel) or Alexa Fluor® 647 Rat Anti-Mouse IL-17 Receptor B (Cat. No. 565866, right panel). Innate lymphoid cells were identified as Lin- CD25+ CD127+ events with the forward and side light-scatter characteristics of viable leukocytes. Approximately 150 events were gated for the generation of contour plots. Flow cytometric analysis was performed using a BD LSRFortessa™ Flow Cytometer System.
Multicolor flow cytometric analysis of IL-17 Receptor B on innate lymphoid cells in mouse bone marrow. BALB/c mouse bone marrow cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) (Cat. No. 553141 or 553142). The cells were then stained with BD Horizon™ V450 Mouse Lineage Antibody Cocktail (Cat. No. 561301), FITC Rat Anti-Mouse CD25 (Cat. No. 553071), PE Rat Anti-Mouse CD127 (Cat. No. 552543), and either Alexa Fluor® 647 Rat IgG1, κ Isotype Control (Cat. No. 557731, left panel) or Alexa Fluor® 647 Rat Anti-Mouse IL-17 Receptor B (Cat. No. 565866, right panel). Innate lymphoid cells were identified as Lin- CD25+ CD127+ events with the forward and side light-scatter characteristics of viable leukocytes. Approximately 150 events were gated for the generation of contour plots. Flow cytometric analysis was performed using a BD LSRFortessa™ Flow Cytometer System.
Preparation And Storage
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- An isotype control should be used at the same concentration as the antibody of interest.
Companion Products
The 6B7 monoclonal antibody specifically binds to IL-17 Receptor B (IL-17RB), which is a single-pass type I transmembrane protein and a component of the heterodimeric receptor for IL-25 (also known as IL-17E). Several components of the IL-17RB signaling mechanism have been identified, including ACT1, TRAF4, SMURF2, DAZAP2, and STAT5. The cytoplasmic tails of both of the IL-25 Receptor components, IL-17RA and IL-17RB, and ACT1 contain the SEFIR (similar expression to fibroblast growth factor genes and IL-17R) domain that is essential for IL-25R-mediated signaling. Signaling through IL-17RB mediates Th2 immune responses and is responsible for pathogenesis in a murine model for asthma. IL-17RB is found on subsets of iNKT cells, Th2 cells, some myeloid cells in solid organs, and type 2 innate lymphoid cells (ILC2) in lung, peritoneum, mesenteric lymph nodes and bone marrow.
Development References (5)
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Hoyler T, Klose CS, Souabni A, et al. The transcription factor GATA-3 controls cell fate and maintenance of type 2 innate lymphoid cells.. Immunity. 2012; 37(4):634-48. (Biology). View Reference
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Terashima A, Watarai H, Inoue S, et al. A novel subset of mouse NKT cells bearing the IL-17 receptor B responds to IL-25 and contributes to airway hyperreactivity.. J Exp Med. 2008; 205(12):2727-33. (Biology). View Reference
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Wu L, Zepp JA, Qian W, et al. A novel IL-25 signaling pathway through STAT5.. J Immunol. 2015; 194(9):4528-34. (Biology). View Reference
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Zepp JA, Wu L, Qian W, et al. TRAF4-SMURF2-mediated DAZAP2 degradation is critical for IL-25 signaling and allergic airway inflammation.. J Immunol. 2015; 194(6):2826-37. (Biology). View Reference
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Zhang B, Liu C, Qian W, Han Y, Li X, Deng J. Crystal structure of IL-17 receptor B SEFIR domain.. J Immunol. 2013; 190(5):2320-6. (Biology). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.
