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Purified Mouse Anti-Human RANTES
Purified Mouse Anti-Human RANTES

Expression of RANTES by cultured human PBMC. Human PBMC were cultured for 24 hours in the presence of BD GolgiStop™ (2 µM final concentration; Cat. No. 554724). The PBMC were harvested, stained with FITC-mouse anti-human CD3 antibody (FITC-HIT3a, Cat. No. 555339), fixed, permeabilized, and subsequently stained with 0.125 µg of PE-mouse anti-human RANTES antibody (PE-2D5, Cat. No. 554732) following BD Biosciences Pharmingen's staining protocol (left panel). To demonstrate specificity of staining, the binding of PE-2D5 antibody was blocked by the preincubation of the unlabeled 2D5 antibody (Cat. No. 556859) with fixed/permeabilized target cells prior to staining with the PE-2D5 antibody. The quadrant markers for the bivariate dot plots were set based on the auto-fluorescence control, and verified with unlabeled antibody blocking control (right panel).

Expression of RANTES by cultured human PBMC. Human PBMC were cultured for 24 hours in the presence of BD GolgiStop™ (2 µM final concentration; Cat. No. 554724). The PBMC were harvested, stained with FITC-mouse anti-human CD3 antibody (FITC-HIT3a, Cat. No. 555339), fixed, permeabilized, and subsequently stained with 0.125 µg of PE-mouse anti-human RANTES antibody (PE-2D5, Cat. No. 554732) following BD Biosciences Pharmingen's staining protocol (left panel). To demonstrate specificity of staining, the binding of PE-2D5 antibody was blocked by the preincubation of the unlabeled 2D5 antibody (Cat. No. 556859) with fixed/permeabilized target cells prior to staining with the PE-2D5 antibody. The quadrant markers for the bivariate dot plots were set based on the auto-fluorescence control, and verified with unlabeled antibody blocking control (right panel).

Product Details
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BD Pharmingen™
Human (QC Testing)
Mouse BALB/c IgG1, κ
Recombinant Human RANTES Protein
Intracellular block/flow cytometry (Routinely Tested), Neutralization, Western blot (Reported)
0.5 mg/ml
AB_398617
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Store undiluted at 4°C.

Recommended Assay Procedures

Recommended Assay Procedure:

Blocking Control for Intracellular Staining: The purified 2D5 antibody (Cat. No. 556859) can be used as a blocking control to demonstrate specificity of human RANTES staining by PE-2D5 antibody (Cat. No. 554732). To perform this control, the fixed/permeabilized cells (~1 million) can be incubated with 1 - 10 µg of unlabeled 2D5 antibody (Cat. No. 556859) for 20 minutes at 4°C, prior to staining with PE-2D5 antibody (eg, 0.1 - 0.5 µg mAb/1 million cells). The intracellular cytokine staining technique and use of blocking controls are described in detail by C. Prussin and D. Metcalfe. For specific methodology, please visit the protocols section or chapter on intracellular staining in the Immune Function Handbook, both of which are posted on our web site, www.bdbiosciences.com.

Neutralization: The 2D5 antibody is useful for neutralization of human RANTES bioactivity. Please note that this application is not routinely tested at BD Biosciences Pharmingen.

WB: The 2D5 antibody has been found useful for use in Western blot. Please note that this application is not routinely tested at BD Biosciences Pharmingen.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  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.
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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.

This antibody is routinely tested by intracellular staining. Other applications were tested at BD Biosciences Pharmingen during antibody development only or reported in the literature.

556859 Rev. 1
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
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Citations & References
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Development References (4)

  1. Leonard EJ, Sylvester I, Yoshimura T, et al. Current Protocols in Immunology. John Wiley and Sons; 1995:6.12.1-6.12.26.
  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: IC/FCM Block). View Reference
  3. 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. (Clone-specific). View Reference
  4. 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. (Clone-specific). View Reference
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556859 Rev. 1

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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.