Clone:
REA693
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MC
Alternative names:
BDCA-2

Extended validation for CD303 (BDCA-2) Antibody, anti-human, REAfinity™

Specificity

Epitope competition
In order to compare the epitope specificity of an antibody, the clone being used is compared with other known clones recognizing the same antigen in a competition assay.
Other clonesOverlap in epitope recognition with REA693
AC144++
201A++
V24-785++
Cells were incubated with an excess of purified unconjugated CD303 (BDCA-2) (REA693) antibody followed by staining with fluorochrome-conjugated antibodies of other known clones against the same marker. Based on the fluorescence signal obtained, the clones were identified as recognizing completely overlapping (++), partially overlapping (+), or completely different epitopes (-) of the marker.

Sensitivity

Performance comparison
Selected fluorochrome conjugated antibodies from Miltenyi Biotec were compared to commercially available hybridoma clones in flow cytometry analysis.
View details
Flow cytometric comparison of different clones for CD303 (BDCA-2). Human peripheral blood mononuclear cells (PBMCs) were stained with CD303 (BDCA-2) antibodies and with a suitable counterstaining. As a control, CD303 (BDCA-2) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and propidium iodide fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD303 (BDCA-2). Human peripheral blood mononuclear cells (PBMCs) were stained with CD303 (BDCA-2) antibodies and with a suitable counterstaining. As a control, CD303 (BDCA-2) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and propidium iodide fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD303 (BDCA-2). Human peripheral blood mononuclear cells (PBMCs) were stained with CD303 (BDCA-2) antibodies and with a suitable counterstaining. As a control, CD303 (BDCA-2) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and propidium iodide fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD303 (BDCA-2). Human peripheral blood mononuclear cells (PBMCs) were stained with CD303 (BDCA-2) antibodies and with a suitable counterstaining. As a control, CD303 (BDCA-2) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and propidium iodide fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Flow cytometric comparison of different clones for CD303 (BDCA-2). Human peripheral blood mononuclear cells (PBMCs) were stained with CD303 (BDCA-2) antibodies and with a suitable counterstaining. As a control, CD303 (BDCA-2) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and propidium iodide fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Fixation data
To provide an indication on how an antibody performs after fixation of cells, in-house data on staining results before and after fixation with 3.7% formaldehyde using Miltenyi Biotec antibodies are provided. Different experimental settings may lead to different results.
No fixation
Post fixation
View details
Comparison of staining pattern on non-fixed and fixed cells using CD303 (BDCA-2) (REA693). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
View details
Comparison of staining pattern on non-fixed and fixed cells using CD303 (BDCA-2) (REA693). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
Comparison of staining pattern on non-fixed and fixed cells using CD303 (BDCA-2) (REA693). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.

Specifications for CD303 (BDCA-2) Antibody, anti-human, REAfinity™

Overview

CD303 (BDCA-2) antibodies have been used, for example, to identify, characterize, and enumerate plasmacytoid dendritic cells in whole blood of healthy and HIV-infected individuals, and for analyzing the role of DC-SIGN in HIV infection and transmission. Furthermore, CD303 (BDCA-2) antibodies have been used to identify and enumerate plasmacytoid dendritic cells in blood and bone marrow samples before and after hematopoietic stem cell mobilization or transplantation. CD303 (BDCA-2) antibodies have also been used for immunohistochemical staining, for example, to identify plasmacytoid dendritic cells in tissue sections from patients with different inflammatory skin diseases.
Clone REA693 recognizes the CD303 (BDCA-2) antigen which is expressed on human plasmacytoid dendritic cells in blood, lymphoid (e.g. tonsils and bone marrow), and non-lymphoid tissue. Specific expression allows direct identification of plasmacytoid dendritic cells using just one marker. CD303 (BDCA-2)
+
plasmacytoid dendritic cells in blood and bone marrow are CD11c
, CD123
high
, CD4
+
, Lin
, CD45RA
+
, CD304 (BDCA-4/Neuropilin-1)
+
, CD141 (BDCA-3)
low
, CD1c (BDCA-1)
, CD14
, and CD2
. They express neither myeloid lineage markers (CD13, CD33) nor Fc receptors (CD32, CD64, FcRI). CD303 (BDCA-2) is strongly expressed on freshly isolated plasmacytoid dendritic cells but down-regulated within 48 hours of culturing. Unlike CD304 (BDCA-4/Neuropilin-1), CD303 (BDCA-2) is not detectable on
ex vivo
generated monocyte-derived or hematopoietic precursor cell-derived CD1a
+
dendritic cells.
The CD303 (BDCA-2) antigen is a novel type II transmembrane C-type lectin. Remarkably, plasmacytoid dendritic cells can take up ligands via CD303 (BDCA-2), then process and present the ligands to T cells. Unlike binding of antibodies to CD304 (BDCA-4), binding of antibodies to CD303 (BDCA-2) inhibits type I IFN production, which is induced in plasmacytoid dendritic cells by, for example, the influenza virus.
Additional information: Clone REA693 displays negligible binding to Fc receptors.

Alternative names

BDCA-2

Detailed product information

Technical specifications

CloneREA693
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman, non-human primate
Cross-reactivity
cynomolgus monkey (
Macaca fascicularis
)
AntigenCD303 (BDCA-2)
Alternative names of antigenBDCA-2
Molecular mass of antigen [kDa]25
Distribution of antigendendritic cells
Entrez Gene ID170482
RRIDAB_2726209, AB_2726484, AB_2726207, AB_2726481, AB_2726204, AB_2819375, AB_2819372, AB_2726485, AB_2726208, AB_2726482, AB_2726205, AB_2726483, AB_2726206, AB_2811648, AB_2726486

Resources for CD303 (BDCA-2) Antibody, anti-human, REAfinity™

Certificates

Please follow this
link
to search for Certificates of Analysis (CoA) by lot number.

References for CD303 (BDCA-2) Antibody, anti-human, REAfinity™

Publications

  1. Contreras, V. et al. (2010) Existence of CD8α-like dendritic cells with a conserved functional specialization and a common molecular signature in distant mammalian species. J. Immunol. 185(6): 3313-3325
  2. Ebner, S. et al. (2002) A novel role for IL-13: Human monocytes cultured in the presence of IL-3 and IL-4 differentiate into dendritic cells that produce less IL-12 and shift the cell responses toward a Tʜ2 cytokine pattern. J. Immunol. 168: 6199-6207
  3. Dzionek, A. et al. (2000) BDCA-2, BDCA-3, BDCA-4: Three markers for distinct subsets of dendritic cells in human peripheral blood. J. Immunol. 165: 6037-6046
  4. Chehimi, J. et al. (2002) Persistent decreases in blood plasmacytoid dendritic cell number and function despite effective highly active antiretroviral therapy and increased blood myeloid dendritic cells in HIV-infected individuals. J. Immunol. 168: 4769-4801
  5. Dutertre, C. A. et al. (2012)
    Pivotal role of M-DC8
    +
    monocytes from viremic HIV-infected patients in TNFα overproduction in response to microbial products.
    Blood 120(11): 2259-2268
  6. De Wit, D. et al. (2004) Blood plasmacytoid dendritic cell responses to CpG oligonucleotides are impaired in human newbornes. Blood 103: 1030-1032
  7. Dzionek, A. et al. (2001) BDCA-2, a novel plasmacytoid dendritic cell–specific type II C-type lectin, mediates antigen capture and is a potent inhibitor of interferon α/β induction. J. Exp. Med. 194: 1823-1834
  8. Rönnblom, L. and Alm, G. V. (2001) A pivotal role for the natural interferon α-producing cells (plasmacytoid dendritic cells) in the pathogenesis of lupus. J. Exp. Med. 194: 59-63
  9. Nestle, F.O. et al. (2005) Plasmacytoid predendritic cells initiate psoriasis through interferon-α production. J. Exp. Med. 202: 135-143
  10. Ebner, S. et al. (2004) Expression of C-type lectin receptors by subsets of dendritic cells in human skin. Int. Immunol. 16: 877-887
  11. Arpinati, M. et al. (2002) Use of anti-BDCA-2 antibody for detection of dendritic cell type-2 (DC2) in allogeneic hematopoietic stem cell transplantation. Bone Marrow Transplant. 29: 887-891
  12. Dzionek, A. et al. (2002) Plasmacytoid dendritic cells: from specific surface markers to specific cellular functions. Hum. Immunol. 63: 1133-1148
  13. Grabbe, S. et al. (2000) Dendritic cells: multi-lineal and multi-functional. Immunol. Today 21: 431-433
  14. Wollenberg, A. et al. (2002) Plasmacytoid dendritic cells: A new cutaneous dendritic cell subset with distinct role in inflammatory skin diseases. J. Invest. Dermatol. 119: 1096-1102
  15. Krupna-Gaylord, M. A. et al. (2014) Induction of type I and type III interferons by Borrelia burgdorferi correlates with pathogenesis and requires linear plasmid 36. PLoS One 9(6): e100174
  16. Clark, F. J. et al. (2003) Origin and subset distribution of peripheral blood dendritic cells in patients with chronic graft-versus-host disease. Transplantation 75: 221-225
  17. Vanders, R. L. et al. (2013) Plasmacytoid dendritic cells and CD8 T cells from pregnant women show altered phenotype and function following H1N1/09 infection. J. Infect. Dis. 208(7): 1062-1070
  18. Zhang, Y. et al. (2013) The influence of cathelicidin LL37 in human anti-neutrophils cytoplasmic antibody (ANCA)-associated vasculitis. Arthritis Res. Ther. 15(5): R161

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