Human DC-SIGN/CD209 Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB161

Clone 120507 was used by HLDA to establish CD designation
R&D Systems, part of Bio-Techne

Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human, Mouse, Rat, Hamster, Primate - Cercopithecus aethiops (African Green Monkey), Primate - Macaca fascicularis (Crab-eating Monkey or Cynomolgus Macaque), Primate - Macaca mulatta (Rhesus Macaque)

Applications

Validated:

Adhesion Blockade, CyTOF-ready, Flow Cytometry, Western Blot

Cited:

Blocking, ELISA Development, Flow Cytometry, Functional Assay, Immunocytochemistry, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Neutralization, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # 120507

View all DC-SIGN/CD209 Antibodies »

Product Specifications

Immunogen

NIH-3T3 mouse embryonic fibroblast cell line transfected with human DC‑SIGN/CD209

Specificity

Detects human DC‑SIGN/CD209 on transfected NIH/3T3 cells and on monocyte derived dendritic cells. Does not react with parental mouse cells or irrelevant transfectants, such as human DC-SIGN2.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Human DC-SIGN/CD209 Antibody

Detection of DC-SIGN antibody in Human DC-SIGN Transfected 3T3 Mouse Cell Line antibody by Flow Cytometry.

Detection of DC-SIGN in Human DC-SIGN Transfected 3T3 Mouse Cell Line by Flow Cytometry.

Human DC-SIGN and DC-SIGN2 transfected 3T3 mouse embryonic fibroblast cell line were stained with Mouse Anti-Human DC-SIGN Monoclonal Antibody (Catalog # MAB161, filled histograms) or isotype control antibody (Catalog # MAB0041, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG F(ab')2Secondary Antibody (Catalog # F0102B).
Detection of DC-SIGN antibody in Human Monocyte Derived Dendritic Cells antibody by Flow Cytometry.

Detection of DC-SIGN in Human Monocyte Derived Dendritic Cells by Flow Cytometry.

Human monocyte derived dendritic cells were stained with Mouse Anti-Human DC-SIGN Monoclonal Antibody (Catalog # MAB161) followed by PE-conjugated anti-mouse IgG (Catalog # F0102B) and Anti-Human B7-2/CD86 Fluorescein-conjugated Monoclonal Antibody (Catalog # FAB141F). Quadrant markers were set based on control antibody staining (Catalog # MAB0041).
Detection of Human DC-SIGN/CD209 by Flow Cytometry

Detection of Human DC-SIGN/CD209 by Flow Cytometry

Infectivity of KSHV is enhanced in the presence of DC-SIGN and DC-SIGNR.A) 293T cells were transfected with empty pcDNA3 vector or expression constructs for DC-SIGN or DC-SIGNR. After 24 hours, cells were infected with 20 µl Bac16 deltaK3 deltaK5 or left uninfected as controls. Cells were harvested after additional 24 hours and surface stained with a DC-SIGN/R antibody (H-200) and analyzed by flow cytometry. Top three panels show transfected cells stained for DC-SIGN/R followed by PE- (DC-SIGN), FITC- (DC-SIGNR) or both (vector) conjugated secondary antibodies. Bottom panels shows KSHV infection of 293T cells transiently expressing DC-SIGN or DC-SIGNR. B, left panels) 293 cell lines stably expressing a vector construct, DC-SIGN or DC-SIGNR were fluorescently stained for surface expression of DC-SIGN or DC-SIGNR. The mean channel fluorescence is indicated in the upper right hand corner. Open histograms – secondary antibody alone; shaded histograms – DC-SIGN or DC-SIGNR staining. B, right panel) 293 pcDNA3, DC-SIGN or DC-SIGNR stable cell lines were pre-incubated with a control antibody (anti-ICAM1, 7 µg/ml), with mannan (100 µg/ml) or a monoclonal antibody specific for DC-SIGN (MAB161; 7 µg/ml) for 30 minutes on ice. These cells were then infected with wild type KSHV (Bac16 or rKSHV.219) at an MOI of 0.01. After 72 hours cells were harvested and evaluated for infection by flow cytometry measuring GFP expression. Infection rates were normalized to 293 pcDNA3 cells treated with the control antibody. The fold increase in relative infectivity is indicated. Data are representative of four independent experiments with two performed in triplicate. C) 293 pcDNA3, DC-SIGN or DC-SIGNR stable lines were infected with 50 µl of concentrated Bac16 wildtype (wt), or mutants with deletion of K3 only ( deltaK3), K5 only ( deltaK5), or deletion of both K3 and K5 ( deltaK3 deltaK5) as indicated. At 72 hours post-infection, the cells were stained for surface expression of DC-SIGN, DC-SIGNR or MHC class I. GFP fluorescence was used as a marker for infection. MHC I staining is shown for infected 293 DC-SIGNR cells. Inset numbers indicate percentage of cells in each quadrant. Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0058056), licensed under a CC-BY license. Not internally tested by R&D Systems.
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Applications for Human DC-SIGN/CD209 Antibody

Application
Recommended Usage

Adhesion Blockade

The adhesion of NIH-3T3 mouse embryonic fibroblast cells (5 x 104 cells/well) to immobilized Recombinant Human ICAM-3/CD50 Fc Chimera (Catalog # 715-IC, 5 µg/mL, 100 µL/well) was maximally inhibited (80-100%) by 5 µg/mL of the antibody.

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

2.5 µg/106 cells
Sample: Human DC-SIGN transfected 3T3 mouse embryonic fibroblast cell line and human monocyte derived dendritic cells

Western Blot

1 µg/mL
Sample: Recombinant Human DC-SIGN Fc Chimera (Catalog # 161-DC)

Reviewed Applications

Read 4 reviews rated 4.3 using MAB161 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Reconstitution Buffer Available:
Size / Price
Qty

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: DC-SIGN/CD209

Human DC-Sign (dendritic cell-specific ICAM-3 grabbing nonintegrin; also CD209) is a member of the chromosome 19 C-type lectin family that includes DC-SIGN, DC-SIGN-related protein, CD23 and LSECtin (1). DC-SIGN was initially reported to be a 46 kDa, 404 amino acid (aa) type II transmembrane protein that contained a 40 aa cytoplasmic N-terminus, a 21 aa transmembrane segment, and a 343 aa extracellular C-terminus (2). The extracellular region contains a distal, 115 aa Ca++-dependent carbohydrate-binding lectin domain and a membrane-proximal linker segment that is composed of seven 23 aa repeats (2, 3). The lectin domain is believed to preferably bind mannose, either within the context of ICAM-3 (on T cells) or ICAM-2 (on endothelial cells) (2, 4, 5). DC-SIGN expression appears to be limited to dendritic cells (DC) and macrophages (6), and DC interaction with the ICAMs both aids DC cell trafficking and immunological synapse formation (7). Since the original report on DC-SIGN, multiple splice forms have been discovered, generating both membrane-bound and soluble forms (3). There are eight type A isoforms, all of which begin with the same 15 aa of exon 1a. Four contain the transmembrane region of exon II, and four do not (i.e., are soluble). Among these eight type A isoforms, only three retain the entire 343 aa found in the full length form described in reference #2 (the full length form is referred to as type I mDC-SIGN1A) (3). Five additional isoforms utilize an alternate start site, and these are referred to as type B isoforms. These all show a 35 aa cytoplasmic domain. One also has a transmembrane segment; four do not. Two of the five contain full, unspliced extracellular regions (3). All of this suggests enormous complexity in DC-SIGN biology. DC-SIGN is not well conserved across species. Human and mouse show little overall aa identity. In the lectin domain, however, human DC-SIGN shares 68% aa identity with mouse DC-SIGN (8). Human and rhesus monkey DC-SIGN share 91% aa identity over the entire extracellular region (8). A detailed description of the additional properties of this monoclonal antibody (MAB161) have been published (9, 10).

References

  1. Liu, W. et al. (2004) J. Biol. Chem. 279:18748.
  2. Curtis, B.M. et al. (1992) Proc. Natl. Acad. Sci. USA 89:8356.
  3. Mummidi, S. et al. (2001) J. Biol. Chem. 276:33196.
  4. Su, S.V. et al. (2004) J. Biol. Chem. 279:19122.
  5. Cambi, A. et al. (2005) Cell. Microbiol. 7:481.
  6. Serrano-Gomez, D. et al. (2004) J. Immunol. 173:5635.
  7. Geijtenbeek, T.B.H. and Y. van Kooyk (2003) Curr. Top. Microbiol. Immunol. 276:32.
  8. Baribaud, F. et al. (2001) J. Virol. 75:10281.
  9. Wu, L. et al. (2002) J. Virol. 76:5905.
  10. Baribaud, F. et al. (2002) J. Virol.76:9135.

Long Name

Dendritic Cell-specific ICAM-3-grabbing Non-integrin 1

Alternate Names

CD209, CLEC4L, DC-SIGN1, DCSIGN

Entrez Gene IDs

30835 (Human); 170786 (Mouse); 102121984 (Cynomolgus Monkey)

Gene Symbol

CD209

Additional DC-SIGN/CD209 Products

Product Documents for Human DC-SIGN/CD209 Antibody

Product Datasheets

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Product Specific Notices for Human DC-SIGN/CD209 Antibody

For research use only

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