Canine CCL2/JE/MCP-1 Biotinylated Antibody

Catalog #: BAF1774 Datasheet / COA / SDS

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BAF1774

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Canine CCL2/JE/MCP-1 Biotinylated Antibody Summary

Species Reactivity

Canine

Specificity

Detects canine CCL2/JE/MCP‑1 in ELISAs and Western blots. In sandwich immunoassay, less than 0.05% cross‑reactivity with recombinant mouse MCP-1 and recombinant human MCP-1 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 BSA as a carrier protein.

Label

Biotin

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

Canine CCL2/JE/MCP-1 Sandwich Immunoassay

Recommended Concentration

Reagent

ELISA Detection (Matched Antibody Pair)

0.1-0.4 µg/mL

Use in combination with:

Capture Reagent: Canine CCL2/JE/MCP‑1 Antibody (Catalog # MAB28171)

Standard: Recombinant Canine CCL2/MCP-1 Protein (Catalog # 1774-MC)

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

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 Goat Anti-Canine CCL2/JE/MCP-1 Biotinylated Antigen Affinity-purified Polyclonal Antibody (Catalog # BAF1774) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Streptavidin (red; Catalog # NL999) and counterstained with DAPI (blue). Specific staining was localized to cytoplasmic. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Shipping

The product is shipped at ambient temperature. Upon receipt, store it 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