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PE-CF594 Mouse Anti-Human CCR9 (CD199)
PE-CF594 Mouse Anti-Human CCR9 (CD199)
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 BD Horizon™ PE-CF594 Mouse IgG1, κ Isotype Control (Cat. No. 562292; Left Plot) or BD Horizon™ PE-CF594 Mouse Anti-Human CCR9 (CD199) antibody (Cat. No. 567982/567983; 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.
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 BD Horizon™ PE-CF594 Mouse IgG1, κ Isotype Control (Cat. No. 562292; Left Plot) or BD Horizon™ PE-CF594 Mouse Anti-Human CCR9 (CD199) antibody (Cat. No. 567982/567983; 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.
Product Details
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BD Horizon™
C-C chemokine receptor type 9; CC-CKR-9; CCR9; CDw199; G protein-coupled receptor 28; GPR-9-6; GPR28; chemokine (C-C motif) receptor 9
Human (QC Testing)
Mouse IgG1, κ
CHO/hCCR9
Flow cytometry (Routinely Tested)
5 µl
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unconjugated antibody and free dye were removed.

Recommended Assay Procedures

BD® CompBeads can be used as surrogates to assess fluorescence spillover (Compensation).  When fluorochrome conjugated antibodies are bound to 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 CompBead to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. 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.
  5. 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).
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. An isotype control should be used at the same concentration as the antibody of interest.
  8. Texas Red is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  9. CF™ is a trademark of Biotium, Inc.
  10. When excited by the yellow-green (561-nm) laser, the fluorescence may be brighter than when excited by the blue (488-nm) laser.
  11. This product is provided under an Agreement between BIOTIUM and BD Biosciences. The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications owned or licensed by Biotium, Inc. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
  12. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using multi-laser cytometers, which may directly excite both PE and CF™594.
  13. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
567982 Rev. 1
Antibody Details
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C9Mab-1

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.

567982 Rev. 1
Format Details
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PE-CF594
BD Horizon™ PE-CF594 dye is a part of the BD PE family of dyes. This tandem fluorochrome is comprised of a R-Phycoerythrin (PE) donor that has excitation maxima (Ex Max) of 496-nm and 566-nm and an acceptor dye with an emission maximum (Em Max) at 615-nm. PE-CF594, driven by BD innovation, is designed to be excited by the blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and detected using an optical filter centered near 615 nm (e.g., a 610/20-nm bandpass filter). The donor dye can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and the acceptor dye can be excited by the green (532-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PE-CF594
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
615 nm
567982 Rev.1
Citations & References
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View product citations for antibody "567982" on CiteAb

Development References (4)

  1. 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
  2. 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
  3. 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
  4. 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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567982 Rev. 1

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.