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 FITC Mouse IgG1, κ Isotype Control (Cat. No. 555748; dashed line histogram) or FITC Mouse Anti-Human CD83 antibody (Cat. No. 556910/560929; 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 cytomtery was performed on a BD FACScan™ system.
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 FITC Mouse IgG1, κ Isotype Control (Cat. No. 555748; dashed line histogram) or FITC Mouse Anti-Human CD83 antibody (Cat. No. 556910/560929; 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 cytomtery was performed on a BD FACScan™ system.
Product Details
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_396534
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 FITC under optimum conditions, and unreacted FITC was removed.
Product Notices
- 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
FITC Mouse IgG1, κ Isotype Control RUO
Size
100 Tests
Cat No.
555748
FITC Mouse Anti-Human CD83 RUO
Size
25 Tests
Cat No.
560929
Recombinant Human IL-4 RUO
Size
5 µg
Cat No.
554605
Recombinant Human GM-CSF RUO
Size
10 µg
Cat No.
550068
556910 Rev. 7
Antibody Details
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.
556910 Rev. 7
Format Details
FITC
Fluorescein (FITC) is part of the BD blue family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 494-nm and an emission maximum (Em Max) at 518-nm. FITC is designed to be excited by the Blue laser (488-nm) and detected using an optical filter centered near 520 nm (e.g., a 530/30-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
FITC
Blue 488 nm
494 nm
518 nm
556910 Rev.7
Citations & References
Development References (8)
-
Hart DN. Dendritic cells: unique leukocyte populations which control the primary immune response. Blood. 1997; 90(9):3245-3287. (Biology). View Reference
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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.
-
Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
-
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.
-
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
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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
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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
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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
556910 Rev. 7
Please refer to Support Documents for Quality Certificates
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.
