Canine CCL2/JE/MCP-1 Antibody

  
  • Species Reactivity
    Canine
  • Specificity
    Detects canine CCL2/JE/MCP-1 in direct ELISAs and Western blots.
  • Source
    Monoclonal Mouse IgG1 Clone # 280702
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    E. coli-derived recombinant canine CCL2/JE/MCP-1
    Gln24-Pro101
    Accession # P52203
  • 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
    <0.10 EU per 1 μg of the antibody by the LAL method.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    Recombinant Canine CCL2/JE/MCP‑1 (Catalog # 1774-MC)
  • Immunocytochemistry
    8-25 µg/mL
    See below
    • Canine CCL2/JE/MCP-1 Sandwich Immunoassay
      Reagent
  • ELISA Capture (Matched Antibody Pair)
    2-8 µg/mL 
    Canine CCL2/JE/MCP‑1 Antibody (Catalog # MAB28171)
  • ELISA Detection (Matched Antibody Pair)
    0.1-0.4 µg/mL 
    Canine CCL2/JE/MCP‑1 Biotinylated Antibody (Catalog # BAF1774)
  • ELISA Standard
     
    Recombinant Canine CCL2/MCP-1 Protein (Catalog # 1774-MC)
  • Neutralization
    Measured by its ability to neutralize CCL2/JE/MCP‑1-induced chemotaxis in the BaF3 mouse pro‑B cell line transfected with human CCR2A. The Neutralization Dose (ND50) is typically 0.5-2.0 µg/mL in the presence of 300 ng/mL Recombinant Canine CCL2/JE/MCP‑1.
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
Chemotaxis Induced by CCL2/JE/MCP‑1 and Neutralization by Canine CCL2/JE/MCP‑1 Antibody. Recombinant Canine CCL2/JE/MCP‑1 (Catalog # 1774-MC) chemoattracts the BaF3 mouse pro‑B cell line transfected with human CCR2A 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 CCL2/JE/MCP‑1 (300 ng/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Canine CCL2/JE/MCP‑1 Monoclonal Antibody (Catalog # MAB28171). The ND50 is typically
0.5-2.0 µg/mL.
Immunocytochemistry
CCL2/JE/MCP‑1 in Canine PBMCs. CCL2/JE/MCP‑1 was detected in immersion fixed canine peripheral blood mononuclear cells (PBMCs) using Mouse Anti-Canine CCL2/JE/MCP‑1 Monoclonal Antibody (Catalog # MAB28171) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 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: CCL2/JE/MCP-1

Canine MCP-1 (monocyte chemotactic protein-1) is an 8 kDa member of the CC chemokine family of chemotactic factors (1, 2). It is synthesized as a 101 amino acid (aa) precursor that contains a 23 aa signal sequence and a 78 aa mature segment (3). It contains no potential N-linked glycosylation sites and is not known for any posttranslational modifications. Based on human studies, MCP-1 will primarily circulate as a monomer. Noncovalent dimers are likely to be found, however. MCP‑1 activity has been localized to the N-terminus (1). Cell types known to secrete MCP-1 are considerable in number, and include keratinocytes, fibroblasts, endothelium, osteoblasts, macrophages, mast cells, smooth muscle cells, and astrocytes (1, 2). In the mature MCP-1 segment, there is 82% and 83% aa identity, canine to human and porcine MCP-1, respectively. When mature canine MCP-1 is compared to (125 aa) extended rodent MCP-1, there is 55% and 56% aa identity, canine to mouse and rat MCP-1, respectively. MCP-1 has three possible receptors. The first two are CCR2 (1) and CCR11 (4). The third receptor has only been identified in mice and is called L-CCR (5). Its function is unknown. MCP-1 is best known as a chemotactic agent for mononuclear cells. It also, however, induces enzyme and cytokine release in monocytes, NK cells, and lymphocytes and histamine release by basophils (1). Additionally, it is believed to reduce IL-12 production by dendritic cells and promote a Th2 phenotype in CD4+ T cells (6).

  • References:
    1. Coillie, E.V. et al. (1999) Cytokine Growth Factor Rev. 10:61.
    2. Yoshie, O. et al. (2001) Adv. Immunol. 78:57.
    3. Kumar, A.G. et al. (1997) Circulation 95:693.
    4. Biber, K. et al. (2003) J. Leukoc. Biol. 74:243.
    5. Luther, S.A. and J.G. Cyster  (2001) Nat. Immunol. 2:102.
  • Entrez Gene IDs:
    6347 (Human); 20296 (Mouse); 24770 (Rat); 403981 (Canine)
  • Alternate Names:
    CCL2; MCAF; MCP-1
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