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Multicolor flow cytometric analysis of CCR9 (CD199) expression on human peripheral blood mononuclear cells. Human PBMCs were stained with BD Horizon™ BUV395 Mouse Anti-Human CD4 antibody (Cat. No. 564724) and APC Mouse IgG1, κ Isotype Control (Cat. No. 554681; Left Plot) or APC Mouse Anti-Human CCR9 (CD199) antibody (Cat. No. 567976/567977; Right Plot). DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. Bivariate pseudocolor density plots showing the correlated expression of CCR9 (CD199) [or Ig Isotype control staining] versus CD4 were derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) lymphocytes. Flow cytometry and data analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
BD Pharmingen™ APC Mouse Anti-Human CCR9 (CD199)
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Recommended Assay Procedures
BD® CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD CompBead to ensure that BD® CompBeads are appropriate for your specific cellular application.
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- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNe, or red diode laser.
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Companion Products
The C9Mab-1 monoclonal antibody specifically binds to Chemokine (C-C motif) receptor 9 (CCR9), which is also known as CD199. CCR9 is an ~42 kDa seven-transmembrane protein in the G protein-coupled receptor (GPCR) supergene family. It is upregulated by CD4+CD8+ double-positive thymocytes, then downregulated in committed CD4+ or CD8+ single-positive thymocytes. CCR9 is also expressed on small intestinal B cells, subsets of memory CD4+ T cells and CD8+ T cells, and dendritic cells. CCR9 binds CCL25, which is also known as thymus-expressed chemokine (TECK) or small inducible chemokine 25 (Scya25). CCL25 is highly expressed in the thymic cortex and small intestinal epithelium. CCR9 is involved in T cell development and gut-associated immune functions via lymphocyte recruitment to the gastrointestinal tract.
Development References (4)
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Olaussen RW, Farstad IN, Brandtzaeg P, Rugtveit J. Age-related changes in CCR9+ circulating lymphocytes: are CCR9+ naive T cells recent thymic emigrants?. Scand J Immunol. 2001; 54(5):435-9. (Biology). View Reference
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Papadakis KA, Prehn J, Nelson V, et al. The role of thymus-expressed chemokine and its receptor CCR9 on lymphocytes in the regional specialization of the mucosal immune system.. J Immunol. 2000; 165(9):5069-76. (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. (Biology). View Reference
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Zabel BA, Agace WW, Campbell JJ, et al. Human G protein-coupled receptor GPR-9-6/CC chemokine receptor 9 is selectively expressed on intestinal homing T lymphocytes, mucosal lymphocytes, and thymocytes and is required for thymus-expressed chemokine-mediated chemotaxis.. J Exp Med. 1999; 190(9):1241-1256. (Biology). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.