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Multicolor flow cytometric analysis of CX3CR1 expression on mouse splenocytes. C57BL/6 mouse splenocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142), then stained with PE Rat Anti-Mouse CD11b (Cat. No. 553311/557397/561689) and either BD Horizon™ BV480 Rat IgG2a, κ Isotype Control (Cat. No. 565630; Left Plot) or BD Horizon™ BV480 Rat Anti-Mouse CX3CR1 antibody (Cat. No. 567824; Right Plot) at 0.25 μg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. Bivariate pseudocolor density plots showing the correlated expression of CX3CR1 versus CD11b (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) cells. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer System. Data shown on this Technical Data Sheet are not lot specific.
BD Horizon™ BV480 Rat Anti-Mouse CX3CR1
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Recommended Assay Procedures
BD® CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
For optimal and reproducible results, BD Horizon Brilliant™ Stain Buffer should be used anytime BD Horizon Brilliant™ dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant™ Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant™ Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant™ Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant™ Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant™ Stain Buffer Plus (Cat. No. 566385).
For Immunofluorescence Applications:
The use of a mounting reagent (eg, ProLong® Gold) is highly recommended to maximize the photostability of BV480. For confocal microscopy systems, a 440 nm laser is the optimal excitation source and the recommended emission filter is a 485/20 nm bandpass filter.
For epifluorescence microscopes with broad spectrum excitation sources, the recommended excitation and emission filters are 445/20 nm and 485/20 nm bandpass filters, respectively. For specific multicolor imaging applications, the exact filter configurations should be optimized by the end user. For additional instrument/filter configuration information, please visit http://www.bdbiosciences.com/research/cellularimaging.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- BD Horizon Brilliant Violet 480 is covered by one or more of the following US patents: 8,575,303; 8,354,239.
- ProLong® is a registered trademark of Thermo Fisher Scientific, Inc. Waltham, MA.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
Companion Products
The Z8-50.23 monoclonal antibody specifically recognizes the mouse CX3C chemokine receptor 1 (CX3CR1), which is also known as Fractalkine receptor, G protein-coupled receptor 13 (GPR13), or GPRV28 (V28). CX3CR1 is an ~40 kDa, seven-transmembrane G protein-coupled receptor that is expressed on monocytes, activated macrophages, NK cells, a subset of memory T cells, dendritic cells (DC), and mast cells. CX3CR1 is a receptor for the CX3C chemokine, CX3CL1, which is also known as fractalkine or neurotactin. CX3CL1 is expressed on activated endothelial cells, neurons, and astrocytes. CX3CR1 plays a role in leucocyte adhesion and extravasation from blood vessels during inflammation.
The antibody was conjugated to BD Horizon™ BV480 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 436-nm and Em Max at 478-nm, BD Horizon™ BV480 can be excited by the violet laser and detected in the BD Horizon™ BV510 (525/40-nm) filter set. BV480 has less spillover into the BV605 detector and, in general, is brighter than BV510.
Development References (4)
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Combadiere C, Gao J, Tiffany HL, Murphy PM. Gene cloning, RNA distribution, and functional expression of mCX3CR1, a mouse chemotactic receptor for the CX3C chemokine fractalkine.. Biochem Biophys Res Commun. 1998; 253(3):728-32. (Biology). View Reference
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Gerlach C, Moseman EA, Loughhead SM, et al. The Chemokine Receptor CX3CR1 Defines Three Antigen-Experienced CD8 T Cell Subsets with Distinct Roles in Immune Surveillance and Homeostasis. Immunity. 2016; 45(6):1270-1284. (Biology). View Reference
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Imai T, Yasuda N. Therapeutic intervention of inflammatory/immune diseases by inhibition of the fractalkine (CX3CL1)-CX3CR1 pathway.. Inflamm Regen. 2016; 36:9. (Biology). View Reference
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Li N, Jiang P, Chen A, et al. CX3CR1 positively regulates BCR signaling coupled with cell metabolism via negatively controlling actin remodeling. Cell Mol Life Sci. 2020; 77(22):4379-4395. (Biology). View Reference
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Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.