Rat CCL4/MIP-1 beta Antibody

  
  • Species Reactivity
    Rat
  • Specificity
    Detects rat CCL4/MIP-1 beta in direct ELISAs and Western blots. In direct ELISAs, approximately 8% cross-reactivity with recombinant mouse CCL4/MIP-1 beta and recombinant rat CCL3/MIP-1 alpha is observed, and less than 2% cross-reactivity with recombinant human CCL4/MIP-1 beta is observed.
  • Source
    Polyclonal Sheep IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant rat CCL4/MIP-1 beta
    Ala24-Asn92
    Accession # P50230
  • 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
  • Neutralization
    Measured by its ability to neutralize CCL4/MIP‑1 beta -induced chemotaxis in the BaF3 mouse proB cell line transfected with human CCR5. The Neutralization Dose (ND50) is typically 0.1-0.5 ug/mL in the presence of 0.075 ug/mL Recombinant rat CCL4/MIP‑1 beta.
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 Rat CCL4/MIP‑1 beta by Western Blot. Western blot shows lysates of NR8383 rat alveolar macrophage cell line untreated (-) or treated (+) with 10 µg/mL LPS for 4 hours. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Rat CCL4/MIP‑1 beta Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6935) 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 1.
Chemotaxis Induced by CCL4/MIP‑1 beta  and Neutralization by Rat CCL4/MIP‑1 beta  Antibody.

Recombinant rat CCL4/MIP‑1 beta (Catalog # 6935-MB) induces BaF3 mouse proB 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 rat CCL4/MIP‑1 beta (0.075 ug/mL) is neutralized (green line) by increasing concentrations of Sheep Anti-Rat CCL4/MIP‑1 beta Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6935). The ND50 is typically 0.1-0.5 ug/mL.

Preparation and Storage
  • Reconstitution
    Sterile PBS to a final concentration of 0.2 mg/mL.
  • 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 inflammation 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 down‑regulation of CCR5 (10). Mature rat CCL4 shares 80% and 86% aa sequence identity with human and mouse CCL4, respectively.
  • 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 Schmid-Grendelmeier, P. (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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