Canine CCL4/MIP-1 beta Antibody

  
  • Species Reactivity
    Canine
  • Specificity
    Detects canine CCL4/MIP‑1 beta in direct ELISAs and Western blots. In direct ELISAs, approximately 10% cross-reactivity with recombinant human CCL4/MIP‑1 beta and recombinant mouse CCL4/MIP‑1 beta is observed and less than 1% cross-reactivity with recombinant viral CCL4/MIP‑1 beta is observed.
  • Source
    Polyclonal Sheep IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant canine CCL4/MIP‑1 beta
    Ala24-Asn92
    Accession # NP_001005250
  • 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.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    See below
  • Immunocytochemistry
    5-15 µg/mL
    Immersion fixed canine peripheral blood mononuclear cells
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 Examples
Detection of Canine CCL4/MIP‑1 beta by Western Blot. Western blot shows lysates of canine peripheral blood mononuclear cells (PBMC) untreated (-) or treated (+) with 5 µg/mL PMA and 500 ng/mL Ionomycin overnight. PVDF Membrane was probed with 1 µg/mL of Canine CCL4/MIP‑1 beta Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5839) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for CCL4/MIP‑1 beta at approximately 12 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • 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: CCL4/MIP-1 beta
CCL4, also known as macrophage inflammatory protein 1 beta (MIP-1 beta, is a 12 kDa beta chemokine that is secreted at sites of inflammation by activated leukocytes, lymphocytes, vascular endothelial cells, and pulmonary smooth muscle cells (1, 2). CCL4 attracts a variety of immune cells to sites of microbial infection as well as to other pathologic inflammations such as allergic asthma and ischemic myocardium (3-8). A CCL4 deficiency in mice promotes the development of autoantibodies, possibly as a result of compromised regulatory T cell recruitment (6). CCL4 is secreted from activated monocytes as a heterodimer with CCL3/MIP-1 alpha (9). The first two N-terminal amino acids can be cleaved from human CCL4 by CD26/DPPIV (10, 11). Both the full length and truncated forms exert biological activity through CCR5, and the truncated form additionally interacts with CCR1 and CCR2 (10). In humans, the ability of CCL4 to bind CCR5 inhibits the cellular entry of M-tropic HIV-1 which utilizes CCR5 as a coreceptor (2). Both forms of CCL4 block HIV-1 infection of T cells by inducing the downregulation of CCR5 (10). Mature canine CCL4 shares 80-84% amino acid sequence identity with human, mouse, and rat CCL4.
  • References:
    1. Rot, A. and U.H. von Andrian (2004) Annu. Rev. Immunol. 22:891.
    2. Menten, P. et al. (2002) Cytokine Growth Factor Rev. 13:455.
    3. Sun, X. et al. (2006) Infec. Immun. 74:5943.
    4. Bisset, L.R. and P. Schmid-Grendelmeier (2005) Curr. Opin. Pulm. Med. 11:35.
    5. Frangogiannis, N.G. (2004) Inflamm. Res. 53:585.
    6. Bystry, R.S. et al. (2001) Nat. Immunol. 2:1126.
    7. Oliveira, S.H.P. et al. (2002) J. Leukoc. Biol. 71:1019.
    8. Schall, T.J. et al. (1993) J. Exp. Med. 177:1821.
    9. Guan, E. et al. (2001) J. Biol. Chem. 276:12404.
    10. Guan, E. et al. (2002) J. Biol. Chem. 277:32348.
    11. Guan, E. et al. (2004) J. Cell. Biochem. 92:53.
  • Entrez Gene IDs:
    6351 (Human); 20303 (Mouse); 116637 (Rat); 448786 (Canine)
  • Alternate Names:
    ACT2; ACT-2; AT744.1; C-C motif chemokine 4; CCL4; chemokine (C-C motif) ligand 4; Exodus-3; G-26 T-lymphocyte-secreted protein; G-26; HC21; LAG1; LAG-1; Lymphocyte activation gene 1 protein; Macrophage inflammatory protein 1-beta; MIP1 beta; MIP-1 beta; MIP1B; MIP1B1; MIP-1-beta; MIP-1-beta(1-69); PAT 744; Protein H400; SCYA2; SCYA4; secreted protein G-26; SIS-gamma; small inducible cytokine A4 (homologous to mouse Mip-1b); Small-inducible cytokine A4; T-cell activation protein 2
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