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BV421 Mouse Anti-Human CX3CL1 (Fractalkine)
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BV421 Mouse Anti-Human CX3CL1 (Fractalkine)
Flow cytometric analysis of CX3CL1 (Fractalkine) expression on Human HepG2 cells.  Cells from the Human HepG2 (Hepatocellular carcinoma, ATCC® HB-8065™) cell line were stained with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat No. 562438, dashed line histogram) or BD OptiBuild™ BV421 Mouse Anti-Human CX3CL1 (Fractalkine) [Cat. No. 752374; solid line histogram] at 1.0 ug/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The fluorescence histogram showing CX3CL1 (Fractalkine) expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) HepG2 cells. Flow cytometry and data analysis were performed using a BD FACSCanto™ II Flow Cytometry System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific
Flow cytometric analysis of CX3CL1 (Fractalkine) expression on Human HepG2 cells.  Cells from the Human HepG2 (Hepatocellular carcinoma, ATCC® HB-8065™) cell line were stained with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat No. 562438, dashed line histogram) or BD OptiBuild™ BV421 Mouse Anti-Human CX3CL1 (Fractalkine) [Cat. No. 752374; solid line histogram] at 1.0 ug/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The fluorescence histogram showing CX3CL1 (Fractalkine) expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) HepG2 cells. Flow cytometry and data analysis were performed using a BD FACSCanto™ II Flow Cytometry System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific
Product Details
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BD OptiBuild™
ABCD-3; C-X3-C motif chemokine 1; CXC3; CXC3C; NTN; NTT; SCYD1; neurotactin
Human (Tested in Development)
Mouse BALB/c IgG1, κ
Human CX3CL1 Recombinant Protein
Flow cytometry (Qualified)
0.2 mg/ml
6376
AB_2875890
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 that minimize unconjugated dye and antibody.

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).

Product Notices

  1. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
  2. Researchers should determine the optimal concentration of this reagent for their individual applications.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  4. 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.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  7. BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
  8. 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.
  9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
752374 Rev. 2
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.

752374 Rev. 2
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV421
Violet 405 nm
407 nm
423 nm
752374 Rev.2
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
752374 Rev. 2

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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.