CCL2/JE/MCP-1 Antibody [Unconjugated]

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.

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.

• Reactivity: Ca
• Applications: WB, B/N, ICC/IF
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

CCL2/JE/MCP-1 Antibody [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant canine CCL2/JE/MCP-1
  Gln24-Pro101
  Accession # P52203
• 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: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: CCL2
• Purity Statement: Antigen Affinity-purified
• Endotoxin Note: <0.10 EU per 1 μg of the antibody by the LAL method.

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 5-15 ug/mL
  • Neutralization 0.8-4.0 ug/mL
  • Western Blot 0.1 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for CCL2/JE/MCP-1 Antibody [Unconjugated]

• C-C motif chemokine ligand 2
• CCL2
• GDCF-2
• HC11
• HSMCR30
• MCAF
• Mcp1
• MCP-1
• SCYA2
• SMC-CF

Background

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).

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for CCL2/JE/MCP-1 Antibody (AF1774)(1)

Publication using AF1774

• DO Riddell, JCW Hildyard, RCM Harron, NL Hornby, DJ Wells, RJ Piercy Serum inflammatory cytokines as disease biomarkers in the DE50-MD dog model of Duchenne muscular dystrophy Disease Models & Mechanisms, 2022-12-09;15(12):. 2022-12-09 [PMID: 36444978] (IHC, Dog)

Bioinformatics

• Gene Symbol: CCL2
• Uniprot:
  • P52203()