CD183 (CXCR3) Antibody, anti-human, REAfinity™

Name: CD183 (CXCR3) Antibody, anti-human, REAfinity™
Availability: InStock

CD183 (CXCR3) Antibody, anti-human, REAfinity™Extended ValidationRecombinant

Clone:

REA232

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC, MICS, IF, IHC, ICC, MC

Alternative names:

CXCR3, CD183, CKR-L2, CMKAR3, GPR9, IP10-R, Mig-R, MigR

Flow cytometry applications forCD183 (CXCR3) Antibody, anti-human, REAfinity™

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

Conjugate:

APC

Recommended dilution:

1:50

Remark:

The antibody is suited for staining of formaldehyde-fixed cells.

Tested:

QC tested

Reported:

31167786,

Products used:

130-120-590, 130-120-450

Protocols:

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

Description:

Human peripheral blood mononuclear cells (PBMCs) were stained with CD183 (CXCR3) antibodies as well as with CD3 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. 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 forCD183 (CXCR3) Antibody, anti-human, REAfinity™

APC
PE

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

PFA

Tested:

during development

Species tested:

human

Products used:

130-120-590, 130-120-450

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:

130-120-590, 130-120-450

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Mass cytometry applications forCD183 (CXCR3) Antibody, anti-human, REAfinity™

pure

Conjugate:

pure

Products used:

130-108-022

Extended validation for CD183 (CXCR3) Antibody, anti-human, REAfinity™

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 REA232
1C6/CXCR3	++
CEW33D	++
G025H7	++
Cells were incubated with an excess of purified unconjugated CD183 (CXCR3) (REA232) 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 CD183 (CXCR3). Human peripheral blood mononuclear cells (PBMCs) were stained with CD183 (CXCR3) antibodies and with a suitable counterstaining. As a control, CD183 (CXCR3) 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 CD183 (CXCR3). Human peripheral blood mononuclear cells (PBMCs) were stained with CD183 (CXCR3) antibodies and with a suitable counterstaining. As a control, CD183 (CXCR3) 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 CD183 (CXCR3). Human peripheral blood mononuclear cells (PBMCs) were stained with CD183 (CXCR3) antibodies and with a suitable counterstaining. As a control, CD183 (CXCR3) 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 CD183 (CXCR3). Human peripheral blood mononuclear cells (PBMCs) were stained with CD183 (CXCR3) antibodies and with a suitable counterstaining. As a control, CD183 (CXCR3) 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 CD183 (CXCR3). Human peripheral blood mononuclear cells (PBMCs) were stained with CD183 (CXCR3) antibodies and with a suitable counterstaining. As a control, CD183 (CXCR3) 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 CD183 (CXCR3) (REA232). 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 CD183 (CXCR3) (REA232). 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 CD183 (CXCR3) Antibody, anti-human, REAfinity™

Overview

Clone REA232 recognizes CD183, a 38 kDa G protein–coupled chemokine receptor. CD183 harbors seven-transmembrane α-helices, an extracellular N-terminal domain containing three loops involved in the chemokine ligand binding, an intracellular C-terminal domain involved in signal transduction. CD183 binds CXCL9, CXCL10, and CXCL11 as ligands. CD183-ligand interactions play role in diverse cellular functions such as chemotactic migration, cell proliferation, and death. Expression of CD183 is found on T cells, human airway epithelial cells, NK cells, plasmacytoid dendritic cells, mast cells, alveolar macrophages, and eosinophils.

Additional information: Clone REA232 displays negligible binding to Fc receptors.

Alternative names

CXCR3, CD183, CKR-L2, CMKAR3, GPR9, IP10-R, Mig-R, MigR

Detailed product information

Technical specifications

Clone	REA232
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	CD183 (CXCR3)
Alternative names of antigen	CXCR3, CD183, CKR-L2, CMKAR3, GPR9, IP10-R, Mig-R, MigR
Distribution of antigen	T cells, NK cells, epithelial cells, mast cells, T cells, NK cells, mast cells, epithelial cells
Entrez Gene ID	2833
RRID	AB_2734058, AB_2751636, AB_2751590, AB_2752156, AB_2752105, AB_2811347, AB_2811334, AB_2784025, AB_2784024, AB_2784023, AB_2784022, AB_2655743, AB_2734057

Resources for CD183 (CXCR3) Antibody, anti-human, REAfinity™

Documents and Protocols

Brochures/posters

REAfinity™ Antibodies for mass cytometry - Recombinantly engineered to increase reproducibility

App notes/customer reports

Comprehensive flow cytometry analysis of chemokine receptors expressed on human natural killer cells.

Certificates

Please follow this

link

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

References for CD183 (CXCR3) Antibody, anti-human, REAfinity™

Publications

Bergmann-Leitner, E. et al. (2018) Identification of immune signatures of novel adjuvant formulations using machine learning. Sci Rep 8(1): 17508
Farooq, F. et al. (2016) Circulating follicular T helper cells and cytokine profile in humans following vaccination with the rVSV-ZEBOV Ebola vaccine. Sci Rep 6: 27944
Hu, J. K. et al. (2011) Expression of chemokine receptor CXCR3 on T cells affects the balance between effector and memory CD8 T-cell generation. Proc. Natl. Acad. Sci. U.S.A. 108: E118-E127
Colvin, R. A. et al. (2004) Intracellular domains of CXCR3 that mediate CXCL9, CXCL10, and CXCL11 function. J. Biol. Chem. 279: 30219-30227

Applications

Extended validation