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BB515 Mouse Anti-Human CX3CL1 (Fractalkine)
BB515 Mouse Anti-Human CX3CL1 (Fractalkine)
Flow cytometric analysis of CX3CL1 expression on HepG2 cells. Human HepG2 cells were stained with either BD Horizon™ BB515 Mouse Anti-Human CX3CL1 antibody (Cat. No. 567748/567749; solid line histogram) or BD Horizon™ BB515 Mouse IgG1, κ Isotype Control (Cat. No. 564416; dashed line histogram). DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. Fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of live (DAPI-negative) cells. Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 Cell Analyzer System and FlowJo™ software.
Flow cytometric analysis of CX3CL1 expression on HepG2 cells. Human HepG2 cells were stained with either BD Horizon™ BB515 Mouse Anti-Human CX3CL1 antibody (Cat. No. 567748/567749; solid line histogram) or BD Horizon™ BB515 Mouse IgG1, κ Isotype Control (Cat. No. 564416; dashed line histogram). DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. Fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of live (DAPI-negative) cells. Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 Cell Analyzer System and FlowJo™ software.
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/immunoassay (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 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 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 CompBead 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 optimal results, it is recommended to perform two washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescence staining, prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.

Product Notices

  1. 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).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. 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.
  7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  8. Alexa Fluor™ is a trademark of Life Technologies Corporation.
567748 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™ BB515 which is part of the BD Horizon Brilliant™ Blue family of dyes. With an Ex Max near 490 nm and an Em Max near 515 nm, BD Horizon™ BB515 can be excited by the blue laser (488 nm) laser and detected with a 530/30 nm filter. This dye has been exclusively developed by BD Biosciences and is up to seven times brighter than FITC with less spillover into the PE channel. Due to similar excitation and emission properties, BB515, FITC, and Alexa Fluor™ 488 cannot be used simultaneously. It is not recommended to use BB515 in cocktails that include Streptavidin conjugates as it may cause high background.

567748 Rev. 1
Format Details
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BB515
The BD Horizon Brilliant™ Blue 515 (BB515) dye is part of the BD Horizon Brilliant™ Blue family of dyes. This dye is a polymer fluorochrome with an excitation maximum (Ex Max) at 490-nm and an emission maximum (Em Max) of 515-nm. Driven by BD innovation, BB515 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 520-nm (e.g., 530/30-nm). BB515 reagents are significantly brighter than equivalent FITC or Alexa Fluor™ 488 reagents with less spillover into the PE detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BB515
Blue 488 nm
490 nm
515 nm
567748 Rev.1
Citations & References
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View product citations for antibody "567748" on CiteAb

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
567748 Rev. 1

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.