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PE-Cy™5 Mouse Anti-Human CD83
PE-Cy™5 Mouse Anti-Human CD83
Flow cytometric analysis of CD83 expression on cultured human dendritic cells. Human peripheral blood monocytes were treated with 20 ng/ml of Recombinant Human IL-4 (Cat. No. 554605), and 20 ng/ml Recombinant Human GM-CSF (Cat. No. 550068) proteins for 7 days at 37°C. In addition, 20 ng/ml Recombinant Human TNF (Cat. No. 554618) was added for the last two days of culture. The cells were then stained with either PE-Cy™5 Mouse IgG1, κ Isotype Control (Cat. No. 555750; dashed line histogram) or PE-Cy™5 Mouse Anti-Human CD83 antibody (Cat. No. 551058; solid line histogram). The fluorescence histogram showing CD83 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable dendritic cells.
Flow cytometric analysis of CD83 expression on cultured human dendritic cells. Human peripheral blood monocytes were treated with 20 ng/ml of Recombinant Human IL-4 (Cat. No. 554605), and 20 ng/ml Recombinant Human GM-CSF (Cat. No. 550068) proteins for 7 days at 37°C. In addition, 20 ng/ml Recombinant Human TNF (Cat. No. 554618) was added for the last two days of culture. The cells were then stained with either PE-Cy™5 Mouse IgG1, κ Isotype Control (Cat. No. 555750; dashed line histogram) or PE-Cy™5 Mouse Anti-Human CD83 antibody (Cat. No. 551058; solid line histogram). The fluorescence histogram showing CD83 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable dendritic cells.
Product Details
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BD Pharmingen™
BL11; HB15; B-cell activation protein
Human (QC Testing), Rhesus, Cynomolgus, Baboon (Tested in Development)
Mouse IgG1, κ
Human CD83 transfected COS cells
Flow cytometry (Routinely Tested)
20 µl
VI
9308
AB_394025
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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. 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.
  7. 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™.
  8. Cy is a trademark of GE Healthcare.
  9. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  10. 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.
  11. 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.
  12. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
551058 Rev. 8
Antibody Details
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HB15e

The HB15e monoclonal antibody specifically binds to a 45 kDa type 1 transmembrane glycoprotein member of the Ig superfamily. CD83 is composed of a single V-type Ig extracellular domain with a C-terminal cytoplasmic tail. Cell surface CD83 is expressed mainly by follicular dendritic cells, circulating dendritic cells, interdigitating dendritic cells in lymphoid tissues, in vitro-generated dendritic cells and thymic dendritic cells. However, its expression is not restricted to dendritic cells. CD83 is also expressed on some germinal center B cells and some lymphoblastoid cell lines. Although its function is not known, it may play a role in cell-cell interaction during antigen presentation.

551058 Rev. 8
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
551058 Rev.8
Citations & References
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Development References (8)

  1. Hart DN. Dendritic cells: unique leukocyte populations which control the primary immune response. Blood. 1997; 90(9):3245-3287. (Biology). View Reference
  2. Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997.
  3. Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
  4. Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995.
  5. Summers KL, Daniel PB, O'Donnell JL, Hart DN. Dendritic cells in synovial fluid of chronic inflammatory arthritis lack CD80 surface expression. Clin Exp Immunol. 1995; 100(1):81-89. (Biology). View Reference
  6. Weissman D, Li Y, Ananworanich J, et al. Three populations of cells with dendritic morphology exist in peripheral blood, only one of which is infectable with human immunodeficiency virus type 1. Proc Natl Acad Sci U S A. 1995; 92(3):826-830. (Biology). View Reference
  7. Zhou LJ, Schwarting R, Smith HM, Tedder TF. A novel cell-surface molecule expressed by human interdigitating reticulum cells, Langerhans cells, and activated lymphocytes is a new member of the Ig superfamily. J Immunol. 1992; 149(2):735-742. (Biology). View Reference
  8. Zhou LJ, Tedder TF. Human blood dendritic cells selectively express CD83, a member of the immunoglobulin superfamily. J Immunol. 1995; 154(8):3821-3835. (Biology). View Reference
View All (8) View Less
551058 Rev. 8

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