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BB700 Mouse Anti-Human CD184
BB700 Mouse Anti-Human CD184

Multiparameter flow cytometric analysis of CD184 expression on human peripheral blood leucocyte populations. Human whole blood was stained with either BD Horizon™ BB700 Mouse IgG2a, κ Isotype Control (Cat. No. 566419; Left Plot) or BD Horizon BB700 Mouse Anti-Human CD184 antibody (Cat. No. 566552/566553; Right Plot). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). A two-parameter flow cytometric dot plot showing the correlated expression of CD184 (CXCR4) [or Ig Isotype control staining] versus side-light scatter (SSC-A) signals was derived from gated events with the forward and side-light scatter characteristics of viable leucocyte populations. Flow cytometric analysis was performed using a BD FACSCelesta™ Flow Cytometer System.

Multiparameter flow cytometric analysis of CD184 expression on human peripheral blood leucocyte populations. Human whole blood was stained with either BD Horizon™ BB700 Mouse IgG2a, κ Isotype Control (Cat. No. 566419; Left Plot) or BD Horizon BB700 Mouse Anti-Human CD184 antibody (Cat. No. 566552/566553; Right Plot). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). A two-parameter flow cytometric dot plot showing the correlated expression of CD184 (CXCR4) [or Ig Isotype control staining] versus side-light scatter (SSC-A) signals was derived from gated events with the forward and side-light scatter characteristics of viable leucocyte populations. Flow cytometric analysis was performed using a BD FACSCelesta™ Flow Cytometer System.

Product Details
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BD Horizon™
CXCR4; Fusin; SDF-1 receptor; LAP3; LCR1; LESTR; NPYY3R; NPY3R; WHIM; HM89
Human (QC Testing), Rhesus, Cynomolgus, Baboon (Tested in Development)
Mouse BALB/c IgG2a, κ
SIVmac variant CP-MAC-infected Sup-T1 cells
Flow cytometry (Routinely Tested)
5 µl
VII 70204, 70305
7852
AB_2869781
Aqueous buffered solution containing ≤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 BD Horizon™ BB700 under optimum conditions, and unconjugated antibody and free BD Horizon BB700 were removed.

Recommended Assay Procedures

     For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions.  More information can be found in the Technical Data Sheet for the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

     When setting up compensation, it is recommended to compare spillover values obtained from cells and BD™ CompBeads to ensure that beads will provide sufficiently accurate spillover values.

     For optimal results, it is recommended to perform two washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescent staining, prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.

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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications. 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. Diagnostic uses require a separate license.
  6. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  7. BD Horizon Brilliant Blue 700 is covered by one or more of the following US patents: 8,455,613 and 8,575,303.
  8. Cy is a trademark of GE Healthcare.
  9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
566552 Rev. 1
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.

The antibody was conjugated to BD Horizon BB700, which is part of the BD Horizon Brilliant™ Blue family of dyes.   It is a polymer-based tandem dye developed exclusively by BD Biosciences.  With an excitation max of 485 nm and an emission max of 693 nm, BD Horizon BB700 can be excited by the 488 nm laser and detected in a standard PerCP-Cy™5.5 set (eg, 695/40-nm filter). This dye provides a much brighter alternative to PerCP-Cy5.5 with less cross laser excitation off the 405 nm and 355 nm lasers.

566552 Rev. 1
Format Details
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BB700
The BD Horizon Brilliant™ Blue 700 (BB700) Dye is part of the BD Horizon Brilliant™ Blue family of dyes. This tandem fluorochrome is comprised of a polymer-technology dye donor with an excitation maximum (Ex Max) of 476-nm and an acceptor dye with an emission maximum (Em Max) at 695-nm. Driven by BD innovation, BB700 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 695-nm (e.g., a 695/20-nm bandpass filter). The donor dye can be excited by the Violet (405 nm) laser and the acceptor dye can be excited by the red (627–640 nm) laser resulting in cross-laser excitation and fluorescence spillover. BB700 Reagents are significantly brighter than equivalent PerCP or PerCP-Cy5.5 reagents and are less sensitive to photobleaching. In addition, BB700 shows much less excitation by the violet (407-nm) laser resulting in less spillover. BB700 has minimal yellow green (562-nm) excitation and is ideal for instruments with both blue (488-nm) and yellow green (562-nm) lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BB700
Blue 488 nm
476 nm
695 nm
566552 Rev.1
Citations & References
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Development References (10)

  1. Bleul CC, Wu L, Hoxie JA, Springer TA, Mackay CR. The HIV coreceptors CXCR4 and CCR5 are differentially expressed and regulated on human T lymphocytes.. Proc Natl Acad Sci U S A. 1997; 94(5):1925-1930. (Clone-specific: Blocking, Flow cytometry, Functional assay, Inhibition, Neutralization). View Reference
  2. 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. (Immunogen: Blocking, Flow cytometry, Fluorescence microscopy, Functional assay, Immunofluorescence, Inhibition, Neutralization). View Reference
  3. 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
  4. Herodin F, Thullier P, Garin D, Drouet M. Nonhuman primates are relevant models for research in hematology, immunology and virology. Eur Cytokine Netw. 2005; 16(2):104-116. (Clone-specific). View Reference
  5. 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
  6. Marcher C, Moller BK, Lillevang ST, Kristensen T. CXCR4 and IL17R are downregulated on cord-blood CD34-positive cells during short-term culture. 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; 2002:629-632.
  7. McKnight A, Wilkinson D, Simmons G, et al. Inhibition of human immunodeficiency virus fusion by a monoclonal antibody to a coreceptor (CXCR4) is both cell type and virus strain dependent. J Virol. 1997; 71(2):1692-1696. (Clone-specific: Blocking, Flow cytometry, Functional assay, Inhibition, Neutralization). View Reference
  8. 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
  9. Vermont-Desroches C, Galea P, Marchand D, Clement C, Wijdenes J. Evaluation of anti-CXCR-4 Antibodies. 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; 2002:247-249.
  10. Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007.
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566552 Rev. 1

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