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Two-color flow cytometric analysis of CCR9 expression on mouse thymocytes and splenocytes. C57BL/6 mouse thymocytes (Top Panels) and splenic leucocytes (Bottom Panels) were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with Rat Anti-Mouse Alexa Fluor® 647 CD8a antibody (Cat No. 557682), and either BD Horizon™ BV421 Mouse IgG2a Isotype Control (Cat. No. 562439; Left Panels) or BD Horizon BV421 Mouse Anti-Mouse CD199 (CCR9) (Right Panels) as indicated. Two-color flow cytometric contour plots showing the correlated expression of CD199 (CCR9) [or Ig Isotype control staining] versus CD8a were derived from gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometric analysis was performed using a BD FACSCanto™ II Flow Cytometer System.


BD Pharmingen™ BV421 Mouse Anti-Mouse CD199 (CCR9)

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- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
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The CW-1.2 monoclonal antibody specifically binds to CD199 which is also known as Chemokine (C-C motif) receptor 9 (CCR9) or C-C chemokine receptor type 9 (C-C CKR-9). CD199 is a ~42 kDa seven-transmembrane protein and member of the G protein-coupled receptor 1 family. CD199 binds to CCL25, which is likewise known as thymus-expressed chemokine (TECK) or small inducible chemokine 25 (Scya25). CCL25 is selectively and constitutively expressed in the thymic cortex and small intestinal epithelium. CD199 is highly expressed on CD4+CD8+ double-positive thymocytes, and mature naïve CD8+ T cells, but not naïve CD4+ T cells, in the peripheral lymphoid organs. CD199 is also expressed on small intestinal B cells, and on subsets of memory CD4+ T cells and CD8+ T cells, and dendritic cells. The CCR9/CCL25 pathway plays important roles in T cell development and gut-associated immune functions. It is especially involved in the recruitment of CD8αα+ intraepithelial lymphocytes (IELs) and the homing of other lymphocytes to the gut. Dysregulation of this pathway is associated with inflammatory responses, such as inflammatory bowel disease (IBD) and celiac disease.
The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.

Development References (7)
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Drakes ML, Stiff PJ, Blanchard TG. Inverse relationship between dendritic cell CCR9 expression and maturation state. Immunology. 2009; 127(4):466-476. (Biology). View Reference
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Feng N, Jaimes MC, Lazarus NH, et al. Redundant role of chemokines CCL25/TECK and CCL28/MEC in IgA+ plasmablast recruitment to the intestinal lamina propria after rotavirus infection. J Immunol. 2006; 176(10):5749-5759. (Biology). View Reference
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Hadeiba H, Lahl K, Edalati A, et al. Plasmacytoid dendritic cells transport peripheral antigens to the thymus to promote central tolerance. Immunity. 2012; 36(3):438-450. (Biology). View Reference
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McGuire HM, Vogelzang A, Ma CS, et al. A subset of interleukin-21+ chemokine receptor CCR9+ T helper cells target accessory organs of the digestive system in autoimmunity. Immunity. 2011; 34(4):602-615. (Biology). View Reference
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Wendland M, Czeloth N, Mach N, et al. CCR9 is a homing receptor for plasmacytoid dendritic cells to the small intestine. Proc Natl Acad Sci U S A. 2007; 104(15):6347-6352. (Biology). View Reference
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Wurbel MA, Malissen B, Campbell JJ. Complex regulation of CCR9 at multiple discrete stages of T cell development. Eur J Immunol. 2006; 36(1):73-81. (Immunogen: Flow cytometry). View Reference
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Wurbel MA, Malissen M, Guy-Grand D, Malissen B, Campbell JJ. Impaired accumulation of antigen-specific CD8 lymphocytes in chemokine CCL25-deficient intestinal epithelium and lamina propria. J Immunol. 2007; 178(12):7598-7606. (Clone-specific: Flow cytometry). View Reference
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