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Flow cytometric analysis of CX3CL1 expression on wild type or Crispr/Cas9 knockout HepG2 cells. Wild type (Left Plot) or knockout (Right Plot) human HepG2 cells were detached with Accutase™ Cell Detachment Solution (Cat. No. 561527) and stained with either BD Horizon™ R718 Mouse Anti-Human CX3CL1 (Fractalkine) antibody (Cat. No. 567762; solid line histograms) or BD Horizon™ R718 Mouse IgG1, κ Isotype Control (Cat. No. 566928; dashed line histograms). DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. Fluorescence histograms showing CX3CL1 expression (or Ig Isotype control) were derived from gated events with the forward and side light-scatter characteristics of live (DAPI-negative) cells. Flow cytometry and data analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Crispr/Cas9 knockout HepG2 was generated by Synthego Corp. Knockout HepG2 cells were bulk sorted and sorted cells were expanded by culturing before staining.
BD Horizon™ R718 Mouse Anti-Human CX3CL1 (Fractalkine)
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- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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The V13-864 monoclonal antibody specifically recognizes CX3C motif chemokine Ligand 1 (CX3CL1), which is also known as fractalkine. CX3CL1 (Fractalkine) is the only known member of the CX3C chemokine subfamily and is expressed as both membrane-bound and secreted forms. Unlike other chemokines, CX3CL1is expressed on the surface of endothelial cells and not significantly expressed on peripheral blood leukocytes. CX3CL1's receptor is known as CX3CR1 and is expressed on cytotoxic T cells, NK cells, dendritic cells, and monocytes. Overexpression of CX3CL1 (Fractalkine) in some malignant tumors may contribute to the recruitment of effectors for innate and adaptive immunity.
The antibody was conjugated to BD Horizon™ Red 718, which has been developed exclusively by for BD Biosciences as a better alternative to Alexa Fluor™ 700. BD Horizon™ Red 718 can be excited by the red laser (628 – 640 nm) and, with an Em Max around 718 nm, it can be detected using a 730/45 nm filter. Due to similar excitation and emission properties, we do not recommend using R718 in combination with APC-R700 or Alexa Fluor™ 700.
Development References (5)
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Bazan JF, Bacon KB, Hardiman G, et al. A new class of membrane-bound chemokine with a CX3C motif.. Nature. 1997; 385(6617):640-4. (Biology). View Reference
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Colobran R, Pujol-Borrell R, Armengol MP, Juan M. The chemokine network. I. How the genomic organization of chemokines contains clues for deciphering their functional complexity.. Clin Exp Immunol. 2007; 148(2):208-17. (Biology). View Reference
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Park MH, Lee JS, Yoon JH. High expression of CX3CL1 by tumor cells correlates with a good prognosis and increased tumor-infiltrating CD8+ T cells, natural killer cells, and dendritic cells in breast carcinoma. J Surg Oncol. 2012; 106(4):386-92. (Biology). View Reference
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Zlotnik A, Yoshie O, Nomiyama H. The chemokine and chemokine receptor superfamilies and their molecular evolution.. Genome Biol. 2006; 7(12):243. (Biology). View Reference
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Zlotnik A, Yoshie O. Chemokines: a new classification system and their role in immunity.. Immunity. 2000; 12(2):121-7. (Biology). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.