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BV421_CD209_overlay.png

Flow cytometric analysis of CD209 expression on peripheral blood monocyte-derived dendritic cells. Adherent peripheral blood mononuclear cells were cultured for 7 days with the recombinant human cytokines, GM-CSF (Cat. No. 550068), TNF (Cat. No. 554618), and IL-4 (Cat. No. 554605). The cultured dendritic cells were harvested and stained with either BD Horizon™ BV421 Mouse IgG2b, κ Isotype Control (Cat. No. 562748, dashed line histogram) or BD Horizon BV421 Mouse Anti-Human CD209 (Cat. No. 564127/566278; solid line histogram). Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

BV421_CD209_overlay.png

Flow cytometric analysis of CD209 expression on peripheral blood monocyte-derived dendritic cells. Adherent peripheral blood mononuclear cells were cultured for 7 days with the recombinant human cytokines, GM-CSF (Cat. No. 550068), TNF (Cat. No. 554618), and IL-4 (Cat. No. 554605). The cultured dendritic cells were harvested and stained with either BD Horizon™ BV421 Mouse IgG2b, κ Isotype Control (Cat. No. 562748, dashed line histogram) or BD Horizon BV421 Mouse Anti-Human CD209 (Cat. No. 564127/566278; solid line histogram). Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

566278.png BV421_CD209_overlay.png BV421_CD209_overlay.png Blue-Cap-Violet-Label.png

Flow cytometric analysis of CD209 expression on peripheral blood monocyte-derived dendritic cells. Adherent peripheral blood mononuclear cells were cultured for 7 days with the recombinant human cytokines, GM-CSF (Cat. No. 550068), TNF (Cat. No. 554618), and IL-4 (Cat. No. 554605). The cultured dendritic cells were harvested and stained with either BD Horizon™ BV421 Mouse IgG2b, κ Isotype Control (Cat. No. 562748, dashed line histogram) or BD Horizon BV421 Mouse Anti-Human CD209 (Cat. No. 564127/566278; solid line histogram). Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

Flow cytometric analysis of CD209 expression on peripheral blood monocyte-derived dendritic cells. Adherent peripheral blood mononuclear cells were cultured for 7 days with the recombinant human cytokines, GM-CSF (Cat. No. 550068), TNF (Cat. No. 554618), and IL-4 (Cat. No. 554605). The cultured dendritic cells were harvested and stained with either BD Horizon™ BV421 Mouse IgG2b, κ Isotype Control (Cat. No. 562748, dashed line histogram) or BD Horizon BV421 Mouse Anti-Human CD209 (Cat. No. 564127/566278; solid line histogram). Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

Product Details
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BD Horizon™
DC-SIGN
Human (QC Testing)
Mouse IgG2b, κ
Human Monocyte Derived DC Cells
Flow cytometry (Routinely Tested)
5 µl
AB_2738610
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 to the dye under optimum conditions and unconjugated antibody and free dye were removed.
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Recommended Assay Procedures

BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation).  When fluorochrome conjugated antibodies are bound to CompBeads, they have spectral properties very similar to cells.   However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls.  It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

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 of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

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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. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
  8. 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.
  9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  10. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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566278 Rev. 3
Antibody Details
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DCN46

The DCN46 antibody specifically binds to dendritic cell-specific ICAM-3 grabbing nonintegrin (DC-SIGN or CD209), a type-II membrane protein of approximately 44 kDa with a mannose-binding C-type lectin domain.  It is highly expressed on dendritic cells in mucosal tissues.  Its sequence is identical to the HIV-1 envelope gp120-binding C-type lectin, and reports suggest that DC-SIGN binds to HIV-1 gp120 and effectively transmits infectious HIV-1 to resting T lymphocytes expressing CD4 and chemokine receptors.  The C-type lectin domain of DC-SIGN is also capable of binding other pathogenic viruses, bacteria, and parasites.  Reports also suggest that DC-SIGN enables the highly efficient migration of dendritic cells from blood into the tissues.  It can interact with ICAM-2, which has a similar sequence as ICAM-3, and is abundantly expressed on vascular and lymphoid endothelium.  Thus, DC-SIGN mediates dendritic cells rolling and transendothelial migration, and its interaction with ICAM-2 is essential to specific migratory functions of dendritic cells.

The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max near 407 nm and Em Max near 421 nm, BD Horizon BV421 can be excited by the violet laser (405 nm) and detected with a 450/50 nm filter. BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific BlueTM conjugates. Due to nearly identical excitation and emission properties but different spillover characteristics, BD Horizon BV421, Pacific Blue, and BD Horizon V450 cannot be used simultaneously.

566278 Rev. 3
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
BV421
407 nm
423 nm
566278 Rev.3
Citations & References
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View product citations for antibody "566278" on CiteAb

Development References (5)

  1. Appelmelk BJ, van Die I, van Vliet SJ, et al. Carbohydrate profiling identifies new pathogens that interact with dendritic cell-specific ICAM-3-grabbing nonintegrin on dendritic cells. J Immunol. 2003; 170(4):1635-1639. (Biology). View Reference
  2. Gruber A, Chalmers AS, Popov S, Ruprecht RM. Functional aspects of binding of monoclonal antibody DCN46 to DC-SIGN on dendritic cells.. Immunol Lett. 2002; 84(2):103-8. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting, Functional assay). View Reference
  3. Sallusto F, Cella M, Danieli C, Lanzavecchia A. Dendritic cells use macropinocytosis and the mannose receptor to concentrate macromolecules in the major histocompatibility complex class II compartment: downregulation by cytokines and bacterial products. J Exp Med. 1995; 182(2):389-400. (Biology). View Reference
  4. Steinman RM, Granelli-Piperno A, Pope M, et al. The interaction of immunodeficiency viruses with dendritic cells. Curr Top Microbiol Immunol. 2003; 276:1-30. (Biology). View Reference
  5. Steinman RM. DC-SIGN: a guide to some mysteries to dendritic cells. Cell. 2000; 100(5):491-494. (Biology). View Reference
View All (5) View Less
566278 Rev. 3

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.