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

Name: CD303 (BDCA-2) Antibody, anti-human, REAfinity™
Availability: InStock

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

Clone:

REA693

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC, MC

Alternative names:

BDCA-2

Flow cytometry applications forCD303 (BDCA-2) Antibody, anti-human, REAfinity™

APC
APC-Vio 770
Biotin
PE
PE-Vio 615
PE-Vio 770
Vio Bright FITC
Vio Bright R720

Conjugate:

APC

Recommended dilution:

1:50

Remark:

The antibody is suited for staining of formaldehyde-fixed cells.

Tested:

QC tested

Products used:

130-114-177, 130-113-651

Protocols:

Cell surface flow cytometry staining protocol (PBS/EDTA/BSA 1:50)

Description:

Human peripheral blood mononuclear cells (PBMCs) were stained with CD303 (BDCA-2) antibodies or with the corresponding REA Control (S) antibodies (left images) as well as with CD123 antibodies. Flow cytometry was performed using the MACSQuant^® Analyzer. The Tandem Signal Enhancer has been used to increase binding specificity of tandem-dye–conjugated antibodies. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence or 4',6-diamidino-2-phenylindole (DAPI) fluorescence, as in the case of tandem conjugates.

Mass cytometry applications forCD303 (BDCA-2) Antibody, anti-human, REAfinity™

pure

Conjugate:

pure

Products used:

130-124-317

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 clones	Overlap 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.

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.

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

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

Clone	REA693
Clonality	monoclonal
Isotype	recombinant human IgG1
Isotype control	REA Control Antibody (S), human IgG1
Host	human cell line
Type of antibody	Primary antibodies, Recombinant antibodies
Species	human, non-human primate
Cross-reactivity	cynomolgus monkey ( Macaca fascicularis )
Antigen	CD303 (BDCA-2)
Alternative names of antigen	BDCA-2
Molecular mass of antigen [kDa]	25
Distribution of antigen	dendritic cells
Entrez Gene ID	170482
RRID	AB_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_2921876, AB_2921864, AB_2726486

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

Documents and Protocols

App notes/customer reports

Clinically applicable CD304 (BDCA-4)⁺ human plasmacytoid dendritic cells – a new era in dendritic cell–based immunotherapy.

App notes/customer reports

Vasoactive intestinal peptide inhibits IFN-α secretion and modulates the immune function of plasmacytoid dendritic cells.

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

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
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
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
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
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
De Wit, D. et al. (2004) Blood plasmacytoid dendritic cell responses to CpG oligonucleotides are impaired in human newbornes. Blood 103: 1030-1032
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
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
Nestle, F. O. et al. (2005) Plasmacytoid predendritic cells initiate psoriasis through interferon-α production. J. Exp. Med. 202: 135-143
Ebner, S. et al. (2004) Expression of C-type lectin receptors by subsets of dendritic cells in human skin. Int. Immunol. 16: 877-887
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
Dzionek, A. et al. (2002) Plasmacytoid dendritic cells: from specific surface markers to specific cellular functions. Hum. Immunol. 63: 1133-1148
Grabbe, S. et al. (2000) Dendritic cells: multi-lineal and multi-functional. Immunol. Today 21: 431-433
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
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
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
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
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

Applications

Extended validation