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BV421 Mouse Anti-Human RANTES
BV421 Mouse Anti-Human RANTES
Two-color flow cytometric analysis of RANTES expression in human peripheral blood lymphocytes. Human peripheral blood mononuclear cells (PBMC) were cultured (5 hr) with Recombinant Human IFN-γ (Cat. No. 554617/554616; 10 ng/ml) and lipopolysaccharide (Sigma, Cat. No. L-8274; 1.0 μg/ml) in the presence of BD GolgiStop™ Protein Transport Inhibitor (containing Monensin) (Cat. No. 554724). The cells were harvested, washed with BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656), and fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655).        The cells were then permeabilized and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with PE Mouse Anti-Human CD3 antibody (Cat. No. 555333/561808/561809) and either BD Horizon™ BV421 Mouse IgG1 κ Isotype Control (Cat. No. 562438; Left Panel) or BD Horizon BV421 Mouse Anti-Human RANTES antibody (Cat. No. 564754; Right Panel) using BD Biosciences Intracellular Cytokine Staining protocol. Two-color flow cytometric contour plots showing the correlated expression patterns of RANTES (or Ig Isotype control staining) versus CD3 were derived from gated events with the forward and side light-scatter characteristics of intact stimulated lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Two-color flow cytometric analysis of RANTES expression in human peripheral blood lymphocytes. Human peripheral blood mononuclear cells (PBMC) were cultured (5 hr) with Recombinant Human IFN-γ (Cat. No. 554617/554616; 10 ng/ml) and lipopolysaccharide (Sigma, Cat. No. L-8274; 1.0 μg/ml) in the presence of BD GolgiStop™ Protein Transport Inhibitor (containing Monensin) (Cat. No. 554724). The cells were harvested, washed with BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656), and fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655).        The cells were then permeabilized and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with PE Mouse Anti-Human CD3 antibody (Cat. No. 555333/561808/561809) and either BD Horizon™ BV421 Mouse IgG1 κ Isotype Control (Cat. No. 562438; Left Panel) or BD Horizon BV421 Mouse Anti-Human RANTES antibody (Cat. No. 564754; Right Panel) using BD Biosciences Intracellular Cytokine Staining protocol. Two-color flow cytometric contour plots showing the correlated expression patterns of RANTES (or Ig Isotype control staining) versus CD3 were derived from gated events with the forward and side light-scatter characteristics of intact stimulated lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Product Details
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BD Horizon™
CCL5; chemokine (C-C motif) ligand 5; EoCP; SCYA5; SIS-delta; SISd; TCP228
Human (QC Testing)
Mouse BALB/c IgG1, κ
Recombinant Human RANTES Protein
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
AB_2738932
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 BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.

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. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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Antibody Details
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2D5

The 2D5 monoclonal antibody specifically binds to human Regulated upon Activation, Normal T cells Expressed and Secreted (RANTES). RANTES belongs to the C-C family of chemokines and is also known as CCL5. RANTES is produced by activated macrophages, CD8+ T cells, platelets, fibroblasts, epithelial cells and some tumor cells. RANTES is a chemoattractant and response modifier for a variety of cells including memory T cells, NK cells, dendritic cells, monocytes, eosinophils, basophils, and mast cells. RANTES binds to and transduces intracellular signals through several cell surface chemokine receptors including CCR1, CCR3, CCR4, and CCR5. RANTES can reportedly inhibit the interaction between certain HIV viruses and CCR5 and thus suppress viral infection in vitro.

The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.

564754 Rev. 1
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
564754 Rev.1
Citations & References
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Development References (5)

  1. Cocchi F, DeVico AL, Garzino-Demo A, Arya SK, Gallo RC, Lusso P. Identification of RANTES, MIP-1 alpha, and MIP-1 beta as the major HIV-suppressive factors produced by CD8+ T cells. Science. 1995; 270(5243):1811-1815. (Clone-specific). View Reference
  2. Prussin C, Metcalfe DD. Detection of intracytoplasmic cytokine using flow cytometry and directly conjugated anti-cytokine antibodies. J Immunol Methods. 1995; 188(1):117-128. (Methodology: Flow cytometry). View Reference
  3. Schall TJ, Bacon K, Toy KJ, Goeddel DV. Selective attraction of monocytes and T lymphocytes of the memory phenotype by cytokine RANTES. Nature. 1990; 347(6294):669-671. (Biology). View Reference
  4. Schall TJ, Jongstra J, Dyer BJ, et al. A human T cell-specific molecule is a member of a new gene family. J Immunol. 1988; 141(3):1018-1025. (Biology). View Reference
  5. Sticherling M, Küpper M, Koltrowitz F, et al. Detection of the chemokine RANTES in cytokine-stimulated human dermal fibroblasts.. J Invest Dermatol. 1995; 105(4):585-91. (Immunogen: Electron microscopy, ELISA, Immunocytochemistry (cytospins), Western blot). View Reference
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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.