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Canine CCL2/JE/MCP-1 Antibody

Catalog #: AF1774
1 Citation Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
AF1774
AF1774-SP
Chemotaxis Induced by CCL2/JE/MCP‑1 and Neutralization by Canine CCL2/JE/MCP‑1 Antibody.
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Canine CCL2/JE/MCP-1 Antibody Summary

Species Reactivity
Canine
Specificity
Detects canine CCL2/JE/MCP-1 in direct ELISAs and Western blots. In direct ELISAs, approximately 40% cross-reactivity with recombinant human (rh) CCL2/MCP-1 is observed, approximately 10% cross-reactivity with recombinant mouse (rm) JE, recombinant rat JE, and recombinant cotton rat JE is observed, and less than 5% cross-reactivity with rhEotaxin, rmMARC, and rhMCP-2 is observed.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
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 either lyophilized or 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
0.1 µg/mL
Recombinant Canine CCL2/JE/MCP‑1 (Catalog # 1774-MC)
Immunocytochemistry
5-15 µg/mL
See below
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 0.8-4.0 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.

Scientific Data

Neutralization Chemotaxis Induced by CCL2/JE/MCP‑1 and Neutralization by Canine CCL2/JE/MCP‑1 Antibody. View Larger

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 Goat Anti-Canine CCL2/JE/ MCP-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1774). The ND50 is typically 0.8-4.0 µg/mL.

Immunocytochemistry CCL2/JE/MCP-1 antibody in Canine PBMCs by Immunocytochemistry (ICC). View Larger

CCL2/JE/MCP‑1 in Canine PBMCs. CCL2/JE/MCP-1 was detected in immersion fixed canine peripheral blood mononuclear cells using Goat Anti-Canine CCL2/JE/MCP-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1774) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
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
C-C motif chemokine ligand 2; CCL2; GDCF-2; HC11; HSMCR30; MCAF; Mcp1; MCP-1; SCYA2; SMC-CF

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Citation for Canine CCL2/JE/MCP-1 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citation: Showing 1 - 1

  1. Serum inflammatory cytokines as disease biomarkers in the DE50-MD dog model of Duchenne muscular dystrophy
    Authors: DO Riddell, JCW Hildyard, RCM Harron, NL Hornby, DJ Wells, RJ Piercy
    Disease Models & Mechanisms, 2022-12-09;15(12):.
    Species: Dog
    Sample Types: Whole Tissue
    Applications: IHC

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