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PerCP-Cy™5.5 Mouse Anti-Human CD184
PerCP-Cy™5.5 Mouse Anti-Human CD184

Flow cytometric analysis of CD184 on Rhesus macaque lysed whole blood.  Rhesus macaque lysed whole blood was stained with the PerCP-Cy™5.5 Mouse Anti-Human CD184 antibody (unshaded) or with a PerCP-Cy™5.5 Mouse IgG2a, κ isotype control (shaded).  Histograms were derived from gated events based on light scattering characteristics for lymphocytes. Flow cytometry was performed on a BD™ LSR II flow cytometry system.

Flow cytometric analysis of CD184 on Rhesus macaque lysed whole blood.  Rhesus macaque lysed whole blood was stained with the PerCP-Cy™5.5 Mouse Anti-Human CD184 antibody (unshaded) or with a PerCP-Cy™5.5 Mouse IgG2a, κ isotype control (shaded).  Histograms were derived from gated events based on light scattering characteristics for lymphocytes. Flow cytometry was performed on a BD™ LSR II flow cytometry system.

Product Details
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BD Pharmingen™
CXCR4; Fusin; SDF-1 receptor; LAP3; LCR1; LESTR; NPYY3R; NPY3R; WHIM; HM89
Rhesus, Cynomolgus, Baboon (QC Testing)
Mouse BALB/c IgG2a, κ
SIVmac variant CP-MAC-infected Sup-T1 cells
Flow cytometry (Routinely Tested)
5 µl
VII 70204, 70305
7852
AB_1727436
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 PerCP-Cy5.5 under optimum conditions, and unconjugated antibody and free PerCP-Cy5.5 were removed. Storage of PerCP-Cy5.5 conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity.

Recommended Assay Procedures

Flow cytometry:  Chemokine receptors are known to internalize during manipulation resulting in low frequency expression.  Investigators are advised to perform immunophenotyping studies of chemokine receptors on freshly collected samples (<24 Hrs). Incubation with the antibody should be done at 4°C in the dark. Cellular manipulation, such as Ficoll separation, freezing, or exposure to cold temperatures prior to staining should be minimized and have been shown to cause a decrease in staining intensity and/or inconsistent results.

Investigators should note that alternative staining procedures may be neccessary.  A multiple-step staining procedure is strongly recommended, in some instances, to amplify immunofluorescent signals for the flow cytometric analysis of CXCR4 expression.  Investigators may find the Purified Mouse Anti-Human CD184 antibody (MN 555972) to be useful in conjunction with appropriate secondary and tertiary reagents for detecting low frequency expression, such as with Biotin Goat Anti-Mouse Ig (MN 553999) and PE Streptavidin (MN 554061) or PerCP-Cy™5.5 Streptavidin (MN 551419).

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. PerCP-Cy5.5–labelled antibodies can be used with FITC- and R-PE–labelled reagents in single-laser flow cytometers with no significant spectral overlap of PerCP-Cy5.5, FITC, and R-PE fluorescence.
  6. PerCP-Cy5.5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using dual-laser cytometers, which may directly excite both PerCP and Cy5.5™. We recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
  7. 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.
  8. This PerCP-conjugated product is sold under license to the following patent: US Patent No. 4,876,190.
  9. Cy is a trademark of Amersham Biosciences Limited.
  10. This product is subject to proprietary rights of Amersham Biosciences Corp. and Carnegie Mellon University and made and sold under license from Amersham Biosciences Corp. This product is licensed for sale only for research. It is not licensed for any other use. If you require a commercial license to use this product and do not have one return this material, unopened to BD Biosciences, 10975 Torreyana Rd, San Diego, CA 92121 and any money paid for the material will be refunded.
  11. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  12. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
560670 Rev. 4
Antibody Details
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12G5

The 12G5 monoclonal antibody specifically binds to CD184, also known as CXCR4 and Fusin. CD184/CXCR4 is a seven-transmembrane domain, G-protein-linked, glycoprotein chemokine receptor. CD184 serves as a receptor for the C-X-C chemokine, SDF-1. It is expressed on a wide variety of hematopoietic cells including lymphoid and myeloid precursor cells, megakaryocytes, platelets, T and B lymphocytes, granulocytes, monocytes/macrophages, and dendritic cells. It is also expressed on vascular endothelial cells, epithelial cells, neurons and astrocytes. CD184 plays a variety of roles in hematopoiesis, vascularization and neural development. CD184 also functions as a coreceptor for infection with T-cell tropic strains of HIV-1 and as a receptor for CD4-independent infection by some HIV isolates. The 12G5 antibody has been reported to block CD4-independent infection by HIV-2 and CD4-dependent infection by some T-cell tropic isolates of HIV-1.

560670 Rev. 4
Format Details
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PerCP-Cy5.5
PerCP-Cy5.5 dye is part of the BD blue family of dyes. This tandem fluorochrome is comprised of a fluorescent protein complex (PerCP) with an excitation maximum (Ex Max) of 482 nm and an acceptor dye with an emission maximum (Em Max) at 676 nm. PerCP-Cy5 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 680 nm (e.g., a 695/40 nm bandpass filter). The donor dye can be partially excited by the Violet (405-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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PerCP-Cy5.5
Blue 488 nm
482 nm
676 nm
560670 Rev.4
Citations & References
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Development References (7)

  1. Endres MJ, Clapham PR, Marsh M, et al. CD4-independent infection by HIV-2 is mediated by fusin/CXCR4. Cell. 1996; 87(4):745-756. (Biology). View Reference
  2. Feng Y, Broder CC, Kennedy PE, Berger EA. HIV-1 entry cofactor: functional cDNA cloning of a seven-transmembrane, G protein-coupled receptor. Science. 1996; 272(5263):872-877. (Biology). View Reference
  3. Guesdon JL, Ternynck T, Avrameas S. The use of avidin-biotin interaction in immunoenzymatic techniques. J Histochem Cytochem. 1979; 27(8):1131-1139. (Biology). View Reference
  4. Loetscher M, Geiser T, O'Reilly T, Zwahlen R, Baggiolini M, Moser B. Cloning of a human seven-transmembrane domain receptor, LESTR, that is highly expressed in leukocytes. J Biol Chem. 1994; 269(1):232-237. (Biology). View Reference
  5. 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.
  6. Nagasawa T, Nakajima T, Tachibana K, et al. Molecular cloning and characterization of a murine pre-B-cell growth-stimulating factor/stromal cell-derived factor 1 receptor, a murine homolog of the human immunodeficiency virus 1 entry coreceptor fusin. Proc Natl Acad Sci U S A. 1996; 93(25):14726-14729. (Biology). View Reference
  7. Simmons G, Wilkinson D, Reeves JD, et al. Primary, syncytium-inducing human immunodeficiency virus type 1 isolates are dual-tropic and most can use either Lestr or CCR5 as coreceptors for virus entry. J Virol. 1996; 70(12):8355-8360. (Biology). View Reference
View All (7) View Less
560670 Rev. 4

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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

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