Canine CCL2/JE/MCP-1 Antibody

Catalog #: MAB28171
1 Citation Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
MAB28171
MAB28171-SP

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

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Canine CCL2/JE/MCP-1 Antibody Summary

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 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

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

Recommended Concentration

Reagent

ELISA Capture (Matched Antibody Pair)

2-8 µg/mL

Use in combination with:

Detection Reagent: Canine CCL2/JE/MCP‑1 Biotinylated Antibody (Catalog # BAF1774)

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.

Scientific Data

Neutralization

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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

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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.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

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

Coillie, E.V. et al. (1999) Cytokine Growth Factor Rev. 10:61.
Yoshie, O. et al. (2001) Adv. Immunol. 78:57.
Kumar, A.G. et al. (1997) Circulation 95:693.
Biber, K. et al. (2003) J. Leukoc. Biol. 74:243.
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

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