Canine CCL5/RANTES Antibody

    
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  • Species Reactivity
    Canine
  • Specificity
    Detects canine CCL5/RANTES in direct ELISAs. In Western blots, no cross-reactivity with recombinant cotton rat CCL5, recombinant feline CCL5, recombinant human (rh) CCL5, 17, 18, recombinant mouse (rm) CCL5, 7, or 18 is observed. In direct ELISAs, approximately 25% cross‑reactivity with recombinant human CCL5 and no cross-reactivity with recombinant CCL5 from cotton rat, cat, or mouse is observed. No cross-reactivity with rhCCL1, 4, 11, 14, 16, 21, 23, 24, 26, 27, rmCCL6, 11, 21, 24, 26, or 27 is observed.
  • Source
    Monoclonal Mouse IgG2B Clone # 676117
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    E. coli-derived recombinant canine CCL5/RANTES
    Ser24-Ser91
    Accession # Q8HYS0
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Endotoxin Level
    <0.01 EU per 1 μg of the antibody by the LAL method.
  • Label
    Unconjugated
Product Datasheets

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Applications
  •  
    Recommended
    Concentration
    Sample
  • Neutralization
    Measured by its ability to neutralize CCL5/RANTES-induced chemotaxis in the BaF3 mouse pro‑B cell line transfected with human CCR5. The Neutralization Dose (ND50) is typically 0.075-0.45 µg/mL in the presence of 10 ng/mL Recombinant Canine CCL5/RANTES.
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Example
Chemotaxis Induced by CCL5/RANTES and Neutralization by Canine CCL5/RANTES Antibody. Recombinant Canine CCL5/RANTES (Catalog # 6045-CR) chemoattracts the BaF3 mouse pro‑B cell line transfected with human CCR5 in a dose-dependent manner (orange line). The amount of cells that migrated through to the lower chemotaxis chamber was measured by Resazurin (Catalog # AR002). Chemotaxis elicited by Recombinant Canine CCL5/RANTES (10 ng/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Canine CCL5/ RANTES Monoclonal Antibody (Catalog # MAB6045). The ND50 is typically 0.075-0.45 µg/mL.
Preparation and Storage
  • Reconstitution
    Sterile PBS to a final concentration of 0.5 mg/mL.
    Reconstitution Buffer Available
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: CCL5/RANTES
CCL5, also known as RANTES (Regulated upon Activation, Normal T cell Expressed and presumably Secreted), is an 8 kDa beta ‑chemokine that plays a primary role in the inflammatory immune response by means of its ability to attract and activate leukocytes (1‑3). Human and mouse RANTES exhibit cross‑species activity on human and mouse cells (4). Mature canine CCL5 shares 74%‑84% amino acid sequence identity with cotton rat, feline, human, mouse, and rat CCL5. CCL5 is secreted by many cell types at inflammatory sites, and it exerts a wide range of activities through the receptors CCR1, CCR3, CCR4, and CCR5 (5, 6). Inflammatory responses can be impaired by the sequestration of CCL5 by the cytomegalovirus protein US28 (7). In humans, CCR5 binding to CCL5 inhibits the infectivity of R5 (M-tropic) but not X4 (T-tropic) strains of HIV-1 (8). The two N-terminal residues of CCL5 can be removed by CD26/DPPIV, generating a protein that functions as a chemotaxis inhibitor and more effectively blocks M-tropic HIV-1 infection of monocytes (9). Oligomerization of CCL5 on glycosaminoglycans is required for CCR1-mediated leukocyte adhesion and activation as well as CCL5’s interaction with the chemokine CXCL4/PF4 (10‑12). The deposition of CCL5 on activated vascular endothelial cells is crucial for monocyte adhesion to damaged vasculature, but CCL5 oligomerization is not required for the extravasation of adherent leukocytes (13‑15). CCL5 is upregulated in breast cancer and promotes tumor progression through the attraction of proinflammatory macrophages in addition to its actions on tumor cells, stromal cells, and the vasculature (16).
  • References:
    1. Schall, T.J. et al. (1990) Nature 347:669.
    2. Bacon, K.B. et al. (1995) Science 269:1727.
    3. Fischer, F.R. et al. (2001) J. Immunol. 167:1637.
    4. Schall, T.J. et al. (1992) Eur. J. Immunol. 22:1477.
    5. Appay, V. and S.L. Rowland-Jones (2001) Trends Immunol. 22:83.
    6. Levy, J.A. (2009) J. Immunol. 182:3945.
    7. Randolph-Habecker, J.R. et al. (2002) Cytokine 19:37.
    8. DeVico, A.L. and Gallo, R.C. (2004) Nat. Rev. Microbiol. 2:401.
    9. Proost, P. et al. (1998) J. Biol. Chem. 273:7222.
    10. Appay, V. et al. (1999) J. Biol. Chem. 274:27505.
    11. Proudfoot, A.E.I. et al. (2003) Proc. Natl. Acad. Sci. 100:1885.
    12. von Hundelshausen, P. et al. (2005) Blood 105:924.
    13. von Hundelshausen, P. et al. (2001) Circulation 103:1772.
    14. Zernecke, A. et al. (2008) Arterioscler. Thromb. Vasc. Biol. 28:1897.
    15. Baltus, T. et al. (2003) Blood 102:1985.
    16. Soria, G. and A. Ben-Baruch (2008) Cancer Lett. 267:271.
  • Entrez Gene IDs:
    6352 (Human); 20304 (Mouse); 403522 (Canine); 493689 (Feline)
  • Alternate Names:
    CCL5; chemokine (C-C motif) ligand 5; D17S136Enormally T-expressed, and presumably secreted; EoCP; Eosinophil chemotactic cytokine; RANTES; SISd; SIS-delta; small inducible cytokine A5 (RANTES); small inducible cytokine subfamily A (Cys-Cys), member 5; Small-inducible cytokine A5; T cell-specific protein P228; T-cell specific protein p288; TCP228T-cell-specific protein RANTES
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Isotype Controls
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Mouse IgG2B Isotype Control

Ctrl MAB004 34  
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Reconstitution Buffers
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Reconstitution Buffer 1 (PBS)

RB01
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Rat Anti-Mouse IgG2B APC-conjugated Antibody

Flow F0133 1  
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Mouse IgG VisUCyte HRP Polymer Antibody

IHC VC001  
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Rat Anti-Mouse IgG2B PE-conjugated Antibody

Flow F0132  
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Mouse F(ab)2 IgG (H+L) PerCP-conjugated Antibody

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Rat Anti-Mouse IgG2B Fluorescein-conjugated Antibody

Flow F0134 1
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Mouse IgG2B Antibody

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