IFNAR2 Antibody, anti-human, REAfinity™

Name: IFNAR2 Antibody, anti-human, REAfinity™
Availability: InStock

IFNAR2 Antibody, anti-human, REAfinity™Extended ValidationRecombinant

Clone:

REA124

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC

Alternative names:

Interferon α/β receptor 2 (IFN-R-2, IFN-α binding protein, IFN-α/β receptor 2), IFN-R, IFN-alpha-REC, IFNABR, IFNARB

Flow cytometry applications forIFNAR2 Antibody, anti-human, REAfinity™

APC
APC-Vio 770
Biotin
PE
PE-Vio 615
PE-Vio 770
Vio Bright V423

Conjugate:

APC

Recommended dilution:

1:11

Remark:

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

Tested:

QC tested

Products used:

130-099-560, 130-099-558

Protocols:

Cell surface flow cytometry staining protocol (PBS/EDTA/BSA 1:11)

Description:

Human peripheral blood mononuclear cells (PBMCs) were stained with Anti-IFNAR2 antibodies or with the corresponding REA Control (S) antibodies (left peak) and analyzed by flow cytometry using the MACSQuant^® Analyzer. CD303 (BDCA-2)⁺ cells were pre-gated for the analysis.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.

Extended validation for IFNAR2 Antibody, anti-human, REAfinity™

Specificity

Other clones	Overlap in epitope recognition with REA124
122	+
Cells were incubated with an excess of purified unconjugated IFNAR2 (REA124) 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.

Knockout validation

To ensure antibody specificity, the target gene is knocked out in a suitable cell line using the CRISPR/Cas9 system and the knockout is confirmed by sequencing of the target locus. The antibody is considered to bind specifically to the intended epitope if no antibody binding to the knockout cells can be detected. The antibody staining is controlled by fluorescence microscopy and/or flow cytometry.

WT

KO

Fluorescence microscopy image of IFNAR2 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with IFNAR2-PE (REA124, red) and counterstained with DRAQ5 (blue) as DNA stain.

Fluorescence microscopy image of IFNAR2 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with IFNAR2-PE (REA124, red) and counterstained with DRAQ5 (blue) as DNA stain.

Overlay histogram showing flow cytometric analysis of human CD45 knockout cells. Wild type (red) and knockout cells (blue) were stained with CD45-PE, clone (REAL258). 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.

Overlay histogram showing flow cytometric analysis of human CD45 knockout cells. Wild type (red) and knockout cells (blue) were stained with CD45-PE, clone (REAL258). 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.

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 IFNAR2. Human peripheral blood mononuclear cells (PBMCs) were stained with IFNAR2 antibodies and with a suitable counterstaining. As a control, IFNAR2 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 IFNAR2. Human peripheral blood mononuclear cells (PBMCs) were stained with IFNAR2 antibodies and with a suitable counterstaining. As a control, IFNAR2 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 IFNAR2. Human peripheral blood mononuclear cells (PBMCs) were stained with IFNAR2 antibodies and with a suitable counterstaining. As a control, IFNAR2 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 IFNAR2 (REA124). 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 IFNAR2 (REA124). 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 IFNAR2 Antibody, anti-human, REAfinity™

Overview

Clone REA124 recognizes the human interferon α/β receptor 2 (IFNAR2) antigen, a 100 kDa single-pass type I membrane protein that forms one of the two chains of a receptor for the type I interferons IFN-α, IFN-β, and IFN-ω. These interferons are a group of structurally and functionally related proteins, induced by viruses or double-stranded RNA and defined by their ability to establish an antiviral state in cells. IFNAR2 is expressed on most lymphocytes, monocytes, and granulocytes, although IFNAR2 expression is higher on monocytes and granulocytes than on lymphocytes. It is involved in IFN-mediated STAT1, STAT2, and STAT3 activation. The alternatively spliced isoform of IFNAR2 is a potent inhibitor of type I IFN receptor activity.

Additional information: Clone REA124 displays negligible binding to Fc receptors.

Alternative names

Interferon α/β receptor 2 (IFN-R-2, IFN-α binding protein, IFN-α/β receptor 2), IFN-R, IFN-alpha-REC, IFNABR, IFNARB

Detailed product information

Technical specifications

Clone	REA124
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
Antigen	IFNAR2
Alternative names of antigen	Interferon α/β receptor 2 (IFN-R-2, IFN-α binding protein, IFN-α/β receptor 2), IFN-R, IFN-alpha-REC, IFNABR, IFNARB
Molecular mass of antigen [kDa]	54,8
Distribution of antigen	lymphocytes, monocytes, fibroblasts
Entrez Gene ID	3455
RRID	AB_2652221, AB_2652222, AB_2652223, AB_2652224, AB_2652225, AB_2652226, AB_2652227, AB_2652228, AB_2652229, AB_2652230, AB_2905263, AB_2905262, AB_2928694, AB_2928693

Resources for IFNAR2 Antibody, anti-human, REAfinity™

Certificates

Please follow this

link

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

References for IFNAR2 Antibody, anti-human, REAfinity™

Publications

Constantinescu, S. N. et al. (1995) Expression and signaling specificity of the IFNAR chain of the type I interferon receptor complex. Proc. Natl. Acad. Sci. U.S.A. 92(23): 10487-10491
Novick, D. et al. (1995) Soluble and membrane-anchored forms of the human IFN-alpha/beta receptor. J. Leukoc. Biol. 57(5): 712-718
Novick, D. et al. (1994) The human interferon alpha/beta receptor: characterization and molecular cloning. Cell 77(3): 391-400

Applications

Extended validation