CD1c (BDCA-1) Antibody, anti-human

Name: CD1c (BDCA-1) Antibody, anti-human
Availability: InStock

CD1c (BDCA-1) Antibody, anti-humanExtended Validation

Clone:

AD5-8E7

Type of antibody:

Primary antibodies

Isotype:

mouse IgG2a

Applications:

FC, MICS, IF, IHC, ICC, MC

Alternative names:

BDCA-1, CD1, R7, M241, T6, CD1a

Flow cytometry applications forCD1c (BDCA-1) Antibody, anti-human

APC
Biotin
FITC
PE
PE-Vio 770
Vio Bright FITC
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:

19171766, 22384272, 22711645, all publications

Products used:

130-113-861, 130-113-299

Protocols:

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

Description:

Human peripheral blood mononuclear cells (PBMCs) were stained with CD1c (BDCA-1) antibodies or with the corresponding isotype control antibodies (left images) as well as with CD19 and CD11c 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. Apart from type-1 myeloid dendritic cells (MDC1s), CD1c (BDCA-1) is also expressed on a subset of B cells. For this reason, B cells were excluded from the analysis according to expression of CD19 and further analysis is restricted to CD19^– cells. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide or 4',6-diamidino-2-phenylindole (DAPI) fluorescence, as in the case of tandem conjugates.

MICS (MACSima Imaging Cyclic Staining) applications forCD1c (BDCA-1) Antibody, anti-human

APC
FITC
PE

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

PFA

Tested:

during development

Species tested:

human

Products used:

130-113-861, 130-113-299

Compatible applications:

IF, IHC, ICC

Protocols:

Protocol for MICS experiments

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

Acetone

Tested:

during development

Species tested:

human

Products used:

130-113-861, 130-113-299

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Mass cytometry applications forCD1c (BDCA-1) Antibody, anti-human

pure

Conjugate:

pure

Products used:

130-108-032

Extended validation for CD1c (BDCA-1) 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 AD5-8E7
REA694	++
L161	-
F10/21A3	-
Cells were incubated with an excess of purified unconjugated CD1c (BDCA-1) (AD5-8E7) 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 CD1c (BDCA-1). Human peripheral blood mononuclear cells (PBMCs) were stained with CD1c (BDCA-1) antibodies and with a suitable counterstaining. As a control, CD1c (BDCA-1) 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 CD1c (BDCA-1). Human peripheral blood mononuclear cells (PBMCs) were stained with CD1c (BDCA-1) antibodies and with a suitable counterstaining. As a control, CD1c (BDCA-1) 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 CD1c (BDCA-1). Human peripheral blood mononuclear cells (PBMCs) were stained with CD1c (BDCA-1) antibodies and with a suitable counterstaining. As a control, CD1c (BDCA-1) 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 CD1c (BDCA-1). Human peripheral blood mononuclear cells (PBMCs) were stained with CD1c (BDCA-1) antibodies and with a suitable counterstaining. As a control, CD1c (BDCA-1) 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 CD1c (BDCA-1). Human peripheral blood mononuclear cells (PBMCs) were stained with CD1c (BDCA-1) antibodies and with a suitable counterstaining. As a control, CD1c (BDCA-1) 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 CD1c (BDCA-1) (AD5-8E7). 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 CD1c (BDCA-1) (AD5-8E7). 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 CD1c (BDCA-1) Antibody, anti-human

Overview

The clone AD5-8E7 recognizes human CD1c, also known as BDCA-1. CD1c is expressed on a major subpopulation of human myeloid dendritic cells (about 0.3% of blood leukocytes). Blood CD1c (BDCA-1)

⁺

myeloid dendritic cells are CD11c

^high

, CD123

^low

, CD4

⁺

, Lin

^–

, CD45RO

⁺

, CD2

⁺

, CD16

^–

, CD141 (BDCA-3)

^low

, CD303 (BDCA-2)

^–

, and CD304 (BDCA-4/Neuropilin-1)

^–

. They express myeloid markers (CD13, CD33) as well as Fc receptors (CD32, CD64, FcεRI) and are of monocytoid appearance. A minor proportion of CD1c (BDCA-1)

⁺

myeloid dendritic cells expresses CD14 and CD11b. In blood, CD1c (BDCA-1) is also expressed on a subpopulation of CD19

⁺

small resting B lymphocytes. CD1c (BDCA-1) expression has also been detected on cortical thymocytes, on Langerhans cells, and on CD1a

⁺

dendritic cells generated

ex vivo

from monocytes or hematopoietic precursor cells. CD1c (BDCA-1)

⁺

myeloid dendritic cells have been designated as type-1 myeloid dendritic cells (MDC1s).

The CD1c antigen is a member of the CD1 family of proteins that are structurally related to MHC class I proteins and mediate the presentation of non-peptide antigens to T cells.

Alternative names

BDCA-1, CD1, R7, M241, T6, CD1a

Detailed product information

Technical specifications

Clone	AD5-8E7
Clonality	monoclonal
Isotype	mouse IgG2a
Isotype control	Isotype Control Antibody, mouse IgG2a
Host	mouse
Type of antibody	Primary antibodies
Species	human, non-human primate
Cross-reactivity	rhesus monkey ( Macaca mulatta ) , cynomolgus monkey ( Macaca fascicularis )
Antigen	CD1c (BDCA-1)
Alternative names of antigen	BDCA-1, CD1, R7, M241, T6, CD1a
Molecular mass of antigen [kDa]	36
Distribution of antigen	B cells, dendritic cells, Langerhans cells, macrophages, monocytes, lymphocytes, T cells, myeloid leukemia cells, thymocytes, B cells, dendritic cells, Langerhans cells, lymphocytes, macrophages, monocytes, myeloid leukemia cells, T cells, thymocytes, B cells, dendritic cells, Langerhans cells, macrophages, monocytes, myeloid leukemia cells, T cells, lymphocytes, thymocytes
Entrez Gene ID	911
RRID	AB_2726080, AB_2726360, AB_2726083, AB_2726358, AB_2726081, AB_2726355, AB_2726078, AB_2726359, AB_2726082, AB_2726356, AB_2726079, AB_2661165, AB_244309, AB_2726357

Resources for CD1c (BDCA-1) Antibody, anti-human

Certificates

Please follow this

link

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

References for CD1c (BDCA-1) Antibody, anti-human

Publications

Carrion, J. et al. (2012) Microbial carriage state of peripheral blood dendritic cells (DCs) in chronic periodontitis influences DC differentiation, atherogenic potential. J. Immunol. 189(6): 3178-3187
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
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
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
Hou, W. et al. (2012) Viral infection triggers rapid differentiation of human blood monocytes into dendritic cells. Blood 119(13): 3128-3131
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
Brodie Miles, E. et al. (2013) Noncanonical dendritic cell differentiation and survival driven by a bacteremic pathogen. J. Leukoc. Biol. 94(2): 281-289
Hémont, C. et al. (2013) Human blood mDC subsets exhibit distinct TLR repertoire and responsiveness. J. Leukoc. Biol. 93(4): 599-609
Brigl, M. and Brenner, M. B. (2004) CD1: antigen presentation and T cell function. Annu. Rev. Immunol. 22: 817-890
Grabbe, S. et al. (2000) Dendritic cells: multi-lineal and multi-functional. Immunol. Today 21: 431-433
Peiser, M. et al. (2003) CD1a and CD1c cell sorting yields a homogeneous population of immature human Langerhans cells. J. Immunol. Methods 279: 41-53
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
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

Applications

Extended validation