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R718 Mouse Anti-Human CX3CL1 (Fractalkine)
R718 Mouse Anti-Human CX3CL1 (Fractalkine)

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.

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.

Product Details
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BD Horizon™
ABCD-3; C-X3-C motif chemokine 1; CXC3; CXC3C; NTN; NTT; SCYD1; neurotactin
Human (QC Testing)
Mouse BALB/c IgG1, κ
Human CX3CL1 Recombinant Protein
Flow cytometry (Routinely Tested)
5 µl
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 unreacted dye was 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 cells and BD CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. 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.
  3. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  4. An isotype control should be used at the same concentration as the antibody of interest.
  5. This product is provided under an Agreement between BIOTIUM and BD Biosciences. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
  6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  7. Alexa Fluor™ is a trademark of Life Technologies Corporation.
  8. Accutase is a registered trademark of Innovative Cell Technologies, Inc.
567762 Rev. 1
Antibody Details
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V13-864

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.

567762 Rev. 1
Format Details
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R718
The BD Horizon™ Red 718 (R718) Dye is part of the BD red family of dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 695-nm and an emission maximum (Em Max) at 718-nm. Driven by BD innovation, R718 is designed to be excited by the red laser (627–640-nm) and detected using an optical filter centered near 720-nm (e.g., a 720/40-nm bandpass filter). R718 is a brighter alternative to Alexa Fluor™ 700. R718 is also a bright small molecule alternative to APC-R700 with lower spread into the APC detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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R718
Red 627-640 nm
695 nm
718 nm
567762 Rev.1
Citations & References
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Development References (5)

  1. 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
  2. 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
  3. 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
  4. 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
  5. Zlotnik A, Yoshie O. Chemokines: a new classification system and their role in immunity.. Immunity. 2000; 12(2):121-7. (Biology). View Reference
View All (5) View Less
567762 Rev. 1

Please refer to Support Documents for Quality Certificates


Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


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.

Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product as described.

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.