CX3CR1 is a 40kD seven transmembrane chemokine receptor that is expressed on several T cell subsets, monocytes, macrophages, microglia and epithelial cells. CX3CR1 binding to membrane bound or soluble CX3CR1/Fractalkine promotes inflammatory responses by inducing monocyte adhesion to endothelial cells and macrophage activation. CX3CR1 polymorphisms are associated with the development of chronic inflammatory disorders. Alternately spliced isoforms have extended N-terminal extracellular regions that increase the potency of CX3CR1 as a fusion coreceptor for HIV-1. Human CX3CR1 shares 82% amino acid sequence identity with mouse and rat CX3CR1.
Applications:
Suitable for Flow Cytometry. Other applications not tested.
Recommended Dilutions:
Flow Cytometry: 10ul labels 1x10e6. Validated using human peripheral blood monocytes.
Optimal dilutions to be determined by the researcher.
Flow Cytometry Protocol:
1. Cells may be Fc-blocked with 1ug of human IgG/10e5 cells for 15 minutes at RT. Do not wash excess blocking IgG from this reaction.
2. Add 10ul of C8299-01F to (same/less than)1x10e6 cells and incubate for 30 minutes at RT.
3. Remove unbound antibody by washing the cells twice in Flow Cytometry Staining Buffer (F4568-10). Note that whole blood requires a RBC lysis step at this point using Flow Cytometry Human Lyse Buffer.
4. Resuspended cells in F4568-10 for flow cytometry analysis. As a control for this analysis, cells in a separate tube should be treated with PE-labeled mouse IgG1 antibody.
Storage and Stability:
May be stored at 4 degrees C before opening. DO NOT FREEZE! Stable at 4 degrees C as an undiluted liquid. Stable for 12 months at 4 degrees C. Freezing R-Phycoerythrin (PE) conjugates will result in a substantial loss of activity. PE conjugates are sensitive to light.
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