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PE-CF594 Rat Anti-Human CX3CR1
PE-CF594 Rat Anti-Human CX3CR1

Multicolor flow cytometric analysis of CX3CR1 expression on human peripheral blood lymphocytes. Whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. After washing, the leucocytes were stained with BD Horizon™ BUV737 Mouse Anti-Human CD56 antibody (Cat. No. 564447; Top Plots), BD Horizon™ BV421 Mouse Anti-Human CD3 antibody (Cat. No. 562426/562427; Bottom Plots), and either BD Horizon™ PE-CF594 Rat IgG2b, κ Isotype Control (Cat. No. 562308; Left Plots) or BD Horizon PE-CF594 Rat Anti-Human CX3CR1 antibody (Cat. No. 565896/565897; Right Plots). Two-color flow cytometric contour plots showing the correlated expression of CD56 or CD3 versus CX3CR1 (or Ig Isotype control staining), were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Flow Cytometer System.

Multicolor flow cytometric analysis of CX3CR1 expression on human peripheral blood lymphocytes. Whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. After washing, the leucocytes were stained with BD Horizon™ BUV737 Mouse Anti-Human CD56 antibody (Cat. No. 564447; Top Plots), BD Horizon™ BV421 Mouse Anti-Human CD3 antibody (Cat. No. 562426/562427; Bottom Plots), and either BD Horizon™ PE-CF594 Rat IgG2b, κ Isotype Control (Cat. No. 562308; Left Plots) or BD Horizon PE-CF594 Rat Anti-Human CX3CR1 antibody (Cat. No. 565896/565897; Right Plots). Two-color flow cytometric contour plots showing the correlated expression of CD56 or CD3 versus CX3CR1 (or Ig Isotype control staining), were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Flow Cytometer System.

Product Details
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BD Horizon™
CCRL1; CMKBRL1; CMKDR1; GPR13; GPRV28; V28; Fractalkine Receptor
Human (QC Testing)
Rat IgG2b, κ
Human CX3CR1 Recombinant Protein
Flow cytometry (Routinely Tested)
5 µl
1524
AB_2869729
Aqueous buffered solution containing BSA and ≤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 with BD Horizon™ PE-CF594 under optimum conditions, and unconjugated antibody and free PE-CF594 were removed.

Recommended Assay Procedures

     For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet for the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

     When setting up compensation, it is recommended to compare spillover values obtained from cells and BD™ CompBeads to ensure that beads will provide sufficiently accurate spillover values.

     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 immunofluorescent 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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  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. When excited by the yellow-green (561-nm) laser, the fluorescence may be brighter than when excited by the blue (488-nm) laser.
  6. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using multi-laser cytometers, which may directly excite both PE and CF™594.
  7. 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.
  8. CF™ is a trademark of Biotium, Inc.
  9. This product is provided under an Agreement between BIOTIUM and BD Biosciences. The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications owned or licensed by Biotium, Inc. 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.
  10. Texas Red is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  11. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  12. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
565897 Rev. 1
Antibody Details
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2A9-1

The 2A9-1 monoclonal antibody specifically binds to human CX3CR1, which is also known as chemokine (C-C) receptor-like 1 (CCRL1), Beta chemokine receptor-like 1 (CMK-BRL-1), G protein-coupled receptor 13 (GPR13), or GPRV28 (V28). CX3CR1 is a seven transmembrane G protein coupled receptor that is expressed by NK cells, T cells, and monocytes. The cellular expression of CX3CR1 is correlated with high levels of intracellular perforin and granzyme B. CX3CR1 serves as a receptor for fractalkine (CX3CL1). Fractalkine is a transmembrane chemokine of the CX3C family that is expressed on activated endothelial cells, neurons, and astrocytes. Interaction of CX3CR1 with fractalkine initiates cellular adhesive and chemotactic responses.

This antibody is conjugated to BD Horizon PE-CF594, which has been developed exclusively by BD Biosciences as a better alternative to PE-Texas Red®. PE-CF594 excites and emits at similar wavelengths to PE-Texas Red® yet exhibits improved brightness and spectral characteristics. Due to PE having maximal absorption peaks at 496 nm and 564 nm, PE-CF594 can be excited by the blue (488-nm), green (532-nm) and yellow-green (561-nm) lasers and can be detected with the same filter set as PE-Texas Red® (eg, 610/20-nm filter).

565897 Rev. 1
Format Details
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PE-CF594
BD Horizon™ PE-CF594 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 with an emission maximum (Em Max) at 615-nm. PE-CF594, driven by BD innovation, is designed to be excited by the blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and detected using an optical filter centered near 615 nm (e.g., a 610/20-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 green (532-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.
altImg
PE-CF594
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
615 nm
565897 Rev.1
Citations & References
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Development References (5)

  1. Kobayashi T, Okamoto S, Iwakami Y, et al. Exclusive increase of CX3CR1+CD28-CD4+ T cells in inflammatory bowel disease and their recruitment as intraepithelial lymphocytes.. Inflamm Bowel Dis. 2007; 13(7):837-46. (Biology). View Reference
  2. Kondo Y, Kimura O, Tanaka Y, et al. Differential Expression of CX3CL1 in Hepatitis B Virus-Replicating Hepatoma Cells Can Affect the Migration Activity of CX3CR1+ Immune Cells.. J Virol. 2015; 89(14):7016-27. (Biology). View Reference
  3. Nanki T, Imai T, Nagasaka K, et al. Migration of CX3CR1-positive T cells producing type 1 cytokines and cytotoxic molecules into the synovium of patients with rheumatoid arthritis.. Arthritis Rheum. 2002; 46(11):2878-83. (Clone-specific: Flow cytometry). View Reference
  4. Nishimura M, Umehara H, Nakayama T, et al. Dual functions of fractalkine/CX3C ligand 1 in trafficking of perforin+/granzyme B+ cytotoxic effector lymphocytes that are defined by CX3CR1 expression.. J Immunol. 2002; 168(12):6173-80. (Immunogen: Flow cytometry). View Reference
  5. Siwetz M, Sundl M, Kolb D, et al. Placental fractalkine mediates adhesion of THP-1 monocytes to villous trophoblast.. Histochem Cell Biol. 2015; 143(6):565-74. (Biology). View Reference
View All (5) View Less
565897 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.