Clone:
REAL222
Type of antibody:
Releasable antibodies, Primary antibodies, Recombinant antibodies
Applications:
FC, MICS, IHC, IF
Alternative names:
NRP1, BDCA-4, NP1, Nrp, VEGF165R, Neuropilin-1

Extended validation for CD304 (BDCA-4) Antibody, anti-human,
REAlease
®

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 REAL222
AD5-17F6++
446921++
U21-1283++
12C2++
REA774++
Cells were incubated with an excess of purified unconjugated CD304 (BDCA-4) (REAL222) 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) (REAL222). 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) (REAL222). 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) (REAL222). 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,
REAlease
®

Overview

Clone REAL222 is an antibody fragment derived from the full CD304 (BDCA-4) antibody molecule. It displays no binding to Fc receptors. The recombinantly engineered antibody fragments are multimerized to form the REAlease Complex to bind markers with high avidity.
Clone REAL222 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 expression, apart from plasmacytoid dendritic cells, is also detected primarily on follicular B helper memory T cells. The 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
+
but CD303 (BDCA-2).
The REAlease Kits consist of the respective fluorochrome-conjugated REAlease Complexes and the REAlease Support Kit for removal of the REAlease Complexes and optional relabeling with different fluorochrome-conjugated REAlease Complexes.

Alternative names

NRP1, BDCA-4, NP1, Nrp, VEGF165R, Neuropilin-1

Detailed product information

Technical specifications

CloneREAL222
Clonalitymonoclonal
Isotype controlControl Antibody
Hostcell line
Type of antibodyReleasable antibodies, Primary antibodies, Recombinant antibodies
Specieshuman
AntigenCD304 (BDCA-4)
Alternative names of antigenNRP1, BDCA-4, NP1, Nrp, VEGF165R, Neuropilin-1
Distribution of antigendendritic cells, B cells, T cells, bone marrow
RRIDAB_2751420, AB_2751271, AB_2751264, AB_2784121, AB_2784120, AB_2751438

Resources for CD304 (BDCA-4) Antibody, anti-human,
REAlease
®

Certificates

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

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

Publications

  1. Grabbe, S. et al. (2000) Dendritic cells: multi-lineal and multi-functional. Immunol. Today 21: 431-433
  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. 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
  4. Dzionek, A. et al. (2002) Plasmacytoid dendritic cells: from specific surface markers to specific cellular functions. Hum. Immunol. 63: 1133-1148
  5. 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
  6. 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
  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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REAlease
®

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