CD183 (CXCR3) Monoclonal Antibody (CXCR3-173), NovaFluor™ Blue 690, eBioscience™

Product Specific Information

Description
The monoclonal antibody CXCR3-173 recognizes mouse CD183 also known as CXCR3. The antibody CXCR3-173 has been shown to affect chemotaxis in response to ligand. The presence of ligand eliminates staining with the antibody. In vivo addition of the antibody delays cardiac and pancreatic islet allograft rejection.

This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.

Applications Tested
This CXCR3-173 antibody has been tested by flow cytometric analysis of mouse splenocytes. This can be used at less than or equal to 0.25 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

Master mixes
• Master mixes of NFs should be made at 2-8 °C and may be made up to 4 hours ahead of time.
• We do not recommend storing master mixes containing NovaFluor conjugates overnight or longer.

Whole Blood compatibility
• When utilizing whole blood (as opposed to density-gradient-purified PBMC), we recommend lysing red blood cells in bulk prior to staining with NovaFluor conjugates.
• See the Bulk Lysis of Human Whole Blood protocol here.
• Staining of whole blood with NovaFluor conjugates followed by lysis of red blood cells may result in higher-than-expected background staining.

Viability dye compatibility
• NovaFluor dyes are not compatible with DNA intercalating viability dyes.
• Do not use viability dyes such as propidium iodide, 7-actinomycin D (7-AAD) and DAPI. Invitrogen LIVE/DEAD Fixable Dead Cell stains are recommended for use with NovaFluor dyes.

CellBlox Plus Blocking Buffer
• This NovaFluor conjugate comes with CellBlox Plus Blocking Buffer (for 25 µg or 25 test products, Cat. No. C001T02F01; for 100 µg or 100 test, Cat. No. C001T03F01), essential for optimal staining.
• Use CellBlox Plus Blocking Buffer in all experiments with NovaFluor conjugates.
• Add 5 µL per sample to antibody cocktails/master mixes (regardless of how many Novafluor-conjugated antibodies are present) before combining with cells.
• CellBlox Plus Blocking Buffer is compatible with either Super Bright Complete Blocking Buffer or Brilliant Stain Buffer and can be used in antibody cocktails/master mixes with those reagents.
• For single-color controls, use 5 µL of CellBlox Plus Blocking Buffer per 100 µL of cell sample (10^3 to 10^8 cells).

NovaFluor conjugates are based on Phiton technology utilizing novel fluorophore-containing nucleic acid dye structures that allow for engineered fluorescent signatures with consideration for spillover and spread impacts. Learn more

Excitation: 494 nm; Emission: 688 nm; Laser: 488 nm (Blue) Laser

Target Information

CD183, also known as CXCR3, is a seven-transmembrane G protein-coupled chemokine receptor that binds CXCL9 (Mig), CXCL10 (IP-10), and CXCL11 (I-TAC), which are part of the CXC chemokine subfamily. CD183 plays a crucial role in leukocyte traffic, influencing integrin activation, cytoskeletal changes, and chemotactic migration. It is expressed on NK cells, subsets of T lymphocytes, regulatory T cells (Tregs), and is preferentially found on Th1-polarized cells. CD183 is prominently expressed in effector/memory T cells and T cells in inflamed tissues, contributing to CD4 T cell responses to grafts, as evidenced by compromised allograft rejection in CXCR3 knockout mice. Chemokine binding induces rapid, short-lived cellular responses due to receptor internalization, with responsiveness restored after receptor recycling. Inhibition by Bordetella pertussis toxin suggests coupling with Gi subclass G proteins. The production of IP-10, Mig, and I-TAC in inflammatory lesions indicates CD183 role in recruiting inflammatory cells, making it a target for developing antagonists to treat inflammatory diseases. Multiple transcript variants encoding different isoforms of CD183 have been identified.