BD Pharmingen™ PE-Cy7 Rat Anti-Mouse CX3CR1
Clone Z8-50 (also known as Z8-50.23) (RUO)
Multicolor flow cytometric analysis of CX3CR1 expressed on mouse splenocytes. Splenic leucocytes from C57BL/6 mice were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with APC Rat Anti-CD11b antibody (Cat. No. 553312/561690) and either PE-Cy7 Mouse IgG2b, κ Isotype Control (Cat. No. 560542, Left Plot) or PE-Cy7 Rat Anti-Mouse CX3CR1 antibody (Cat. No. 567820, Right Plot) at 0.5 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The bivariate pseudocolor density plot showing CX3CR1 versus CD11b expression (or Ig Isotype control staining) was derived from gated events with the forward and side-light scatter characteristics of viable (7-AAD-negative) leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
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Multicolor flow cytometric analysis of CX3CR1 expressed on mouse splenocytes. Splenic leucocytes from C57BL/6 mice were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with APC Rat Anti-CD11b antibody (Cat. No. 553312/561690) and either PE-Cy7 Mouse IgG2b, κ Isotype Control (Cat. No. 560542, Left Plot) or PE-Cy7 Rat Anti-Mouse CX3CR1 antibody (Cat. No. 567820, Right Plot) at 0.5 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The bivariate pseudocolor density plot showing CX3CR1 versus CD11b expression (or Ig Isotype control staining) was derived from gated events with the forward and side-light scatter characteristics of viable (7-AAD-negative) leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
BD Pharmingen™
CCRL1; CMKBRL1; CMKDR1; Chemokine (C-X3-C motif) receptor 1; Fractalkine receptor; GPR13; V28
Mouse (QC Testing)
Rat F344, also known as Fischer, CDF IgG2a, κ
Mouse CX3CR1 Transfected Cell Line
Flow cytometry (Routinely Tested)
0.2 mg/ml
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unconjugated antibody and free dye were removed.
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 cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Warning: Some APC-Cy7 and PE-Cy7 conjugates show changes in their emission spectrum with prolonged exposure to formaldehyde. If you are unable to analyze fixed samples within four hours, we recommend that you use BD™ Stabilizing Fixative (Cat. No. 338036).
- An isotype control should be used at the same concentration as the antibody of interest.
- PE-Cy7 is a tandem fluorochrome composed of R-phycoerythrin (PE), which is excited by 488-nm light and serves as an energy donor, coupled to the cyanine dye Cy7, which acts as an energy acceptor and fluoresces maximally at 780 nm. PE-Cy7 tandem fluorochrome emission is collected in a detector for fluorescence wavelengths of 750 nm and higher. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from PE may be observed. Therefore, we recommend that individual compensation controls be performed for every PE-Cy7 conjugate. PE-Cy7 is optimized for use with a single argon ion laser emitting 488-nm light, and there is no significant overlap between PE-Cy7 and FITC emission spectra. When using dual-laser cytometers, which may directly excite both PE and Cy7, we recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
Size
0.5 mg
Cat No.
553142
APC Rat Anti-CD11b RUO
Size
0.1 mg
Cat No.
553312
PE-Cy™7 Mouse IgG2b, κ Isotype Control RUO
Size
0.1 mg
Cat No.
560542
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567820 Rev. 1
Antibody Details
Z8-50
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.
567820 Rev. 1
Format Details
PE-Cy7
PE-Cy7 dye is a part of the BD PE family of dyes. This tandem fluorochrome is comprised of a R-Phycoerythrin (PE) donor that has excitation maxima (Ex Max) of 496-nm and 566-nm and an acceptor dye, Cy™7, with an emission maximum (Em Max) at 781-nm. PE can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and detected using an optical filter centered near 781 nm (e.g., a 760/60-nm bandpass filter). The donor dye can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and the acceptor dye can be excited by the Red (627–640-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
PE-Cy7
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
781 nm
567820 Rev.1
Citations & References
View product citations for antibody "567820" on CiteAb
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
567820 Rev. 1
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.