Clone:
AD5-17F6
Type of antibody:
Primary antibodies
Isotype:
mouse IgG1
Applications:
FC, MICS, IF, IHC, MC
Alternative names:
NRP1, BDCA-4, NP1, Nrp, VEGF165R

Extended validation for CD304 (BDCA-4) 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 clonesOverlap in epitope recognition with AD5-17F6
REA774++
446921++
U21-1283++
12C2++
REAL222++
Cells were incubated with an excess of purified unconjugated CD304 (BDCA-4) (AD5-17F6) 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 CD304 (BDCA-4). Human peripheral blood mononuclear cells (PBMCs) were stained with CD304 (BDCA-4) antibodies and with a suitable counterstaining. As a control, CD304 (BDCA-4) 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.
View details
Flow cytometric comparison of different clones for CD304 (BDCA-4). Human peripheral blood mononuclear cells (PBMCs) were stained with CD304 (BDCA-4) antibodies and with a suitable counterstaining. As a control, CD304 (BDCA-4) 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.
View details
Flow cytometric comparison of different clones for CD304 (BDCA-4). Human peripheral blood mononuclear cells (PBMCs) were stained with CD304 (BDCA-4) antibodies and with a suitable counterstaining. As a control, CD304 (BDCA-4) 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.
View details
Flow cytometric comparison of different clones for CD304 (BDCA-4). Human peripheral blood mononuclear cells (PBMCs) were stained with CD304 (BDCA-4) antibodies and with a suitable counterstaining. As a control, CD304 (BDCA-4) 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.
View details
Flow cytometric comparison of different clones for CD304 (BDCA-4). Human peripheral blood mononuclear cells (PBMCs) were stained with CD304 (BDCA-4) antibodies and with a suitable counterstaining. As a control, CD304 (BDCA-4) 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.
View details
Flow cytometric comparison of different clones for CD304 (BDCA-4). Human peripheral blood mononuclear cells (PBMCs) were stained with CD304 (BDCA-4) antibodies and with a suitable counterstaining. As a control, CD304 (BDCA-4) 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 CD304 (BDCA-4). Human peripheral blood mononuclear cells (PBMCs) were stained with CD304 (BDCA-4) antibodies and with a suitable counterstaining. As a control, CD304 (BDCA-4) 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
View details
Comparison of staining pattern on non-fixed and fixed cells using CD304 (BDCA-4) (AD5-17F6). 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 CD304 (BDCA-4) (AD5-17F6). 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 CD304 (BDCA-4) (AD5-17F6). 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 CD304 (BDCA-4) Antibody, anti-human

Overview

Clone AD5-17F6 recognizes the human CD304 (BDCA-4) antigen. Like CD303 (BDCA-2), the CD304 (BDCA-4) antigen is specifically expressed on human plasmacytoid dendritic cells in blood and bone marrow. Unlike CD303 (BDCA-2), expression of CD304 (BDCA-4) is up-regulated on plasmacytoid dendritic cells and induced on myeloid cells upon culturing. In inflamed tonsils, CD304 (BDCA-4) expression, apart from plasmacytoid dendritic cells, is also detected primarily on follicular B helper memory T cells. CD304 (BDCA-4) is identical to neuropilin-1 (NP-1), a neuronal receptor, which is known to be expressed on numerous nonhematopoietic cell types, including neurons, endothelial, and tumor cells. CD1a
+
dendritic cells generated
ex vivo
from monocytes or hematopoietic precursor cells are CD304 (BDCA-4)
+
but CD303 (BDCA-2)
.

Alternative names

NRP1, BDCA-4, NP1, Nrp, VEGF165R

Detailed product information

Technical specifications

CloneAD5-17F6
Clonalitymonoclonal
Isotypemouse IgG1
Isotype controlIsotype Control Antibody, mouse IgG1
Hostmouse
Type of antibodyPrimary antibodies
Specieshuman
AntigenCD304 (BDCA-4)
Alternative names of antigenNRP1, BDCA-4, NP1, Nrp, VEGF165R
Molecular mass of antigen [kDa]101
Distribution of antigenchondrocytes, dendritic cells, endothelial cells, neurons, T cells, CNS cells, heart, kidney, liver, placenta, chondrocytes, dendritic cells, endothelial cells, neurons, T cells, heart, kidney, liver, placenta
Entrez Gene ID8829
RRIDAB_2734056, AB_2751190, AB_2751124, AB_2733316, AB_2733317, AB_2733186, AB_2733187, AB_2751189, AB_2751123, AB_2661197, AB_244174, AB_2734055

References for CD304 (BDCA-4) Antibody, anti-human

Publications

  1. 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
  2. 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
  3. 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
  4. 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
  5. Dzionek, A. et al. (2002) Plasmacytoid dendritic cells: from specific surface markers to specific cellular functions. Hum. Immunol. 36: 1133-1148
  6. Grabbe, S. et al. (2000) Dendritic cells: multi-lineal and multi-functional. Immunol. Today 21: 431-433
  7. 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

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