CD141 (BDCA-3) Antibody, anti-human

Name: CD141 (BDCA-3) Antibody, anti-human
Availability: InStock

CD141 (BDCA-3) Antibody, anti-humanExtended Validation

Clone:

AD5-14H12

Type of antibody:

Primary antibodies

Isotype:

mouse IgG1κ, mouse IgG1

Applications:

FC, MICS, IF, IHC, MC

Alternative names:

THBD, AHUS6, BDCA3, THPH12, THRM, TM

Flow cytometry applications forCD141 (BDCA-3) Antibody, anti-human

APC
APC-Vio 770
Biotin
FITC
PE
PE-Vio 770
Vio Bright FITC
VioBlue
pure

Conjugate:

APC

Recommended dilution:

1:50

Remark:

Cells should be stained prior to fixation, if formaldehyde is used as a fixative.

Tested:

QC tested

Reported:

20479118, 22611244, 22711645, all publications

Products used:

130-113-876, 130-113-314

Protocols:

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

Description:

Human peripheral blood mononuclear cells (PBMCs) were stained with CD141 (BDCA-3) antibodies as well as with Anti-CLEC9A and CD45 antibodies and analyzed by flow cytometry using the MACSQuant^® Analyzer. The Tandem Signal Enhancer has been used to increase binding specificity of tandem-dye–conjugated antibodies. For all other conjugates the FcR Blocking Reagent has been used to avoid Fc receptor–mediated antibody labeling. A pre-gate of CD45⁺ cells was used. 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.

MICS (MACSima Imaging Cyclic Staining) applications forCD141 (BDCA-3) Antibody, anti-human

APC
FITC
PE

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

PFA

Tested:

during development

Species tested:

human

Products used:

130-113-876, 130-113-314

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

Acetone

Tested:

during development

Species tested:

human

Products used:

130-113-876, 130-113-314

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Mass cytometry applications forCD141 (BDCA-3) Antibody, anti-human

pure

Conjugate:

pure

Products used:

130-108-033

Extended validation for CD141 (BDCA-3) Antibody, anti-human

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 AD5-14H12
501733	-
1A4	-
REA674	++
JAA17	-
M80	-
Cells were incubated with an excess of purified unconjugated CD141 (BDCA-3) (AD5-14H12) 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 CD141 (BDCA-3). Human peripheral blood mononuclear cells (PBMCs) were stained with CD141 (BDCA-3) antibodies and with a suitable counterstaining. As a control, CD141 (BDCA-3) 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 4',6-diamidino-2-phenylindole (DAPI) 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 CD141 (BDCA-3). Human peripheral blood mononuclear cells (PBMCs) were stained with CD141 (BDCA-3) antibodies and with a suitable counterstaining. As a control, CD141 (BDCA-3) 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 4',6-diamidino-2-phenylindole (DAPI) 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 CD141 (BDCA-3). Human peripheral blood mononuclear cells (PBMCs) were stained with CD141 (BDCA-3) antibodies and with a suitable counterstaining. As a control, CD141 (BDCA-3) 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 4',6-diamidino-2-phenylindole (DAPI) 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 CD141 (BDCA-3). Human peripheral blood mononuclear cells (PBMCs) were stained with CD141 (BDCA-3) antibodies and with a suitable counterstaining. As a control, CD141 (BDCA-3) 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 4',6-diamidino-2-phenylindole (DAPI) 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 CD141 (BDCA-3). Human peripheral blood mononuclear cells (PBMCs) were stained with CD141 (BDCA-3) antibodies and with a suitable counterstaining. As a control, CD141 (BDCA-3) 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 4',6-diamidino-2-phenylindole (DAPI) 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 CD141 (BDCA-3) (AD5-14H12). 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 CD141 (BDCA-3) (AD5-14H12). 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 CD141 (BDCA-3) Antibody, anti-human

Overview

Clone AD5-14H12 recognizes the human CD141 (BDCA-3) antigen which is expressed at high levels on a minor subpopulation of human myeloid dendritic cells (about 0.02% of blood leukocytes). CD141 (BDCA‑3)

^high

blood dendritic cells are CD11c

^dim

, CD123

^–

, CD4

⁺

, Lin

^–

, CD45RO

⁺

, CD2

^–

, and CD16

^–

. They express myeloid lineage markers, such as CD13 and CD33, and have a monocytoid morphology. Unlike CD1c (BDCA-1)

⁺

blood dendritic cells, CD141 (BDCA-3)

^high

blood dendritic cells lack expression of CD2 and Fc receptors such as CD32, CD64, or FcεRI. CD141 (BDCA-3) is also present at very low levels on CD14

⁺

monocytes, granulocytes, CD303 (BDCA‑2)

⁺

CD304 (BDCA-4/Neuropilin-1)

⁺

plasmacytoid and CD1c (BDCA-1)

⁺

myeloid dendritic cells. CD141 (BDCA‑3)

^high

CD1c (BDCA-1)

^–

myeloid dendritic cells have been designated type-2 myeloid dendritic cells (MDC2s). CD141 is also known as thrombomodulin; thrombomodulin mediates co-agglutination by interaction with thrombin and protein C, though nothing is known about its function on MDC2s.

Alternative names

THBD, AHUS6, BDCA3, THPH12, THRM, TM

Detailed product information

Technical specifications

Clone	AD5-14H12
Clonality	monoclonal
Isotype	mouse IgG1κ, mouse IgG1
Isotype control	Isotype Control Antibody, mouse IgG1
Host	mouse
Type of antibody	Primary antibodies
Species	human
Antigen	CD141 (BDCA-3)
Alternative names of antigen	THBD, AHUS6, BDCA3, THPH12, THRM, TM
Molecular mass of antigen [kDa]	59
Distribution of antigen	endothelial cells, megakaryocytes, monocytes, platelets, neutrophils, smooth muscle, endothelial cells, megakaryocytes, monocytes, platelets, myeloid dendritic cells, neutrophils, smooth muscle
Entrez Gene ID	7056
RRID	AB_2726094, AB_2726375, AB_2726098, AB_2726372, AB_2726095, AB_2733312, AB_2733313, AB_2726374, AB_2726097, AB_2726373, AB_2726096, AB_2726369, AB_2726092, AB_2726370, AB_2726093, AB_244169, AB_2661189, AB_2726371

Resources for CD141 (BDCA-3) Antibody, anti-human

Certificates

Please follow this

link

to search for Certificates of Analysis (CoA) by lot number.

References for CD141 (BDCA-3) Antibody, anti-human

Publications

Gosh, M. et al. (2012) CD13 regulates dendritic cell cross-presentation and T cell responses by inhibiting receptor-mediated antigen uptake. J Immunol 188(11): 5489-5499
Mittag, D. et al. (2011) Human dendritic cell subsets from spleen and blood are similar in phenotype and function but modified by donor health status. J Immunol 186(11): 6207-6217
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
Bosco, M. C. et al. (2011)
Hypoxia modulates the gene expression profile of immunoregulatory receptors in human mature dendritic cells: identification of TREM-1 as a novel hypoxic marker
in vitro
and
in vivo
.
Blood 117(9): 2625-2639
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
van de Ven, R. et al. (2011) Characterization of four conventional dendritic cell subsets in human skin-draining lymph nodes in relation to T-cell activation. Blood 118(9): 2502-2510
MacDonald, K. P. A. et al. (2002) Characterization of human blood dendritic cell subsets. Blood 100: 4512-4520
Chu, C. C. et al. (2012)
Resident CD141 (BDCA3)
⁺
dendritic cells in human skin produce IL-10 and induce regulatory T cells that suppress skin inflammation.
J. Exp. Med. 209(5): 935-945
Jongbloed, S. L. et al. (2010)
Human CD141
⁺
(BDCA-3)
⁺
dendritic cells (DCs) represent a unique myeloid DC subset that cross-presents necrotic cell antigens.
J. Exp. Med. 207(6): 1247-1260
Hémont, C. et al. (2013) Human blood mDC subsets exhibit distinct TLR repertoire and responsiveness. J. Leukoc. Biol. 93(4): 599-609
Lebre, C. M. et al. (2003)
BDCA-3
^hi
dendritic cells: a novel subset with distinct phenotypical characteristics. Doctoral dissertation.
University of Amsterdam, Netherlands
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
Jardine, L. et al. (2013) Rapid detection of dendritic cell and monocyte disorders using CD4 as a lineage marker of the human peripheral blood antigen-presenting cell compartment. Front Immunol 4: 495
Oosterhoff, D. et al. (2012) Tumor-mediated inhibition of human dendritic cell differentiation and function is consistently counteracted by combined p38 MAPK and STAT3 inhibition. Oncoimmunology 1(5): 649-658

Applications

Extended validation