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PE-Cy™5 Mouse Anti-Human CD183
PE-Cy™5 Mouse Anti-Human CD183

Flow cytometric analysis of CD183 expression on human peripheral blood lymphocytes. Human whole blood was stained with either PE-Cy™5 Mouse Anti-Human CD183 (Cat. No. 561731/551128; solid line histogram) or PE-Cy™5 Mouse IgG1 κ Isotype Control (Cat. No. 561731; dashed line histogram). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Fluorescent histograms were derived from gated events with the side and forward light-scattering characteristics of viable lymphocytes.

Flow cytometric analysis of CD183 expression on human peripheral blood lymphocytes. Human whole blood was stained with either PE-Cy™5 Mouse Anti-Human CD183 (Cat. No. 561731/551128; solid line histogram) or PE-Cy™5 Mouse IgG1 κ Isotype Control (Cat. No. 561731; dashed line histogram). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Fluorescent histograms were derived from gated events with the side and forward light-scattering characteristics of viable lymphocytes.

Product Details
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BD Pharmingen™
CXCR3; C-X-C chemokine receptor type 3; GPR9; IP10R; MigR; CKRL2; CMKAR3
Human (QC Testing), Rhesus, Cynomolgus, Baboon (Tested in Development)
Mouse BALB/c IgG1, κ
Human CXCR3 Peptide
Flow cytometry (Routinely Tested)
20 µl
VII 70500
2833
AB_394061
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 with PE-Cy5 (formerly known as BD Cy-Chrome™) under optimum conditions, and unconjugated antibody and free PE-Cy5 were removed.

Product Notices

  1. 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).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  5. 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.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. PE-Cy5 is a tandem fluorochrome composed of R-phycoerythrin (PE), which is excited by the 488 nm light of an Argon ion laser and serves as an energy donor, coupled to the cyanine dye Cy5, which acts as an energy acceptor and fluoresces at 670 nm. BD Biosciences Pharmingen has maximized the fluorochrome energy transfer in PE-Cy5, thus maximizing its fluorescence emission intensity, minimizing residual emission from PE, and minimizing lot-to-lot variation.
  8. PE-Cy5 tandem fluorochromes have been reported to bind some classes of human macrophages and granulocytes via Fc receptors, and PE has been reported to bind to mouse B lymphocytes via Fc receptors. Preincubation of mouse leukocytes with Mouse BD Fc Block™ purified anti-mouse CD16/CD32 mAb 2.4G2 can reduce the non-specific binding of PE-Cy5-conjugated reagents to mouse B cells. However, PE-Cy5 conjugated reagents should not be used to stain splenocytes of SJL, NOD, and MRL mice as B lymphocytes and/or other leukocytes have been reported to non-specifically stain regardless of the use of Mouse BD Fc Block™ (the CD72c complex has been implicated for PE-Cy5 binding in these strains). Reagents conjugated to PE, PerCP, PerCP-Cy5.5, APC, and APC-Cy7 tandem fluorochrome can be used on leukocytes from these mouse strains.
  9. PE-Cy5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the PE-Cy5 tandem fluorochrome, extra care must be taken when using dual-laser cytometers which may directly excite both PE and Cy5™.
  10. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  11. Cy is a trademark of GE Healthcare.
  12. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
551128 Rev. 5
Antibody Details
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1C6/CXCR3

The 1C6/CXCR3 monoclonal antibody specifically binds to human CD183, also known as the CXCR3 chemokine receptor. CD183 is a 40-41 kDa seven-transmembrane protein and member of the G protein-coupled receptor family. CD183 is expressed primarily on activated T cells that infiltrate inflammatory sites. It has also been detected on some circulating T cells, B cells, and NK cells. Reports show that some CXCR3-positive T cells also express CCR5 and are mostly CD45RO-positive cells. Three ligands for CXCR3 have been identified. They are CXCL9 (Mig/monokine induced by interferon-γ), CXCL10 (IP-10/interferon-γ inducible 10-kD protein), and CXCL11 (I-TAC/interferon-inducible T-cell alpha chemoattractant). These chemokines are produced by a variety of cells upon stimulation by IFN-γ and interact with CXCR3 to mediate T-cell chemotaxis. This reagent has been reported to be suitable for immunohistochemical staining of acetone-fixed, frozen sections and/or formalin-fixed, paraffin-embedded tissue sections with citrate pretreatment. Clone 1C6/CXCR3 also cross reacts with a subset of peripheral blood lymphocytes of baboon, and both rhesus and cynomolgus macaque monkeys. The distribution of lymphocytes is similar to that observed with CD183-positive peripheral blood lymphocytes from normal human donors. CXCR3 has been clustered as CD183 in the VIIth HLDA workshop.  

551128 Rev. 5
Format Details
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PE-Cy5
PE-Cy5 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, Cy™5, with an emission maximum (Em Max) at 670-nm. PE 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 670-nm (e.g., a 670/20-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 Red (627-640-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-Cy5
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
670 nm
551128 Rev.5
Citations & References
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Development References (4)

  1. Loetscher M, Gerber B, Loetscher P, et al. Chemokine receptor specific for IP10 and mig: structure, function, and expression in activated T-lymphocytes. J Exp Med. 1996; 184(3):963-969. (Biology). View Reference
  2. Piali L, Weber C, LaRosa G, et al. The chemokine receptor CXCR3 mediates rapid and shear-resistant adhesion-induction of effector T lymphocytes by the chemokines IP10 and Mig. Eur J Immunol. 1998; 28(3):961-972. (Biology). View Reference
  3. Qin S, Rottman JB, Myers P, et al. The chemokine receptors CXCR3 and CCR5 mark subsets of T cells associated with certain inflammatory reactions. J Clin Invest. 1998; 101(4):746-754. (Clone-specific). View Reference
  4. Uguccioni M, Willimann K. Cytokine/Chemokine Receptors: Section report. In: Mason D. David Mason .. et al., ed. Leucocyte typing VII : white cell differentiation antigens : proceedings of the Seventh International Workshop and Conference held in Harrogate, United Kingdom. Oxford: Oxford University Press; 2002:237-243.
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551128 Rev. 5

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