Equine IL-1ra/IL-1F3 Antibody

Catalog #: AF2466 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF2466
AF2466-SP

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Equine IL-1ra/IL-1F3 Antibody Summary

Species Reactivity

Equine

Specificity

Detects equine IL‑1ra/IL‑1F3 in ELISAs and Western blots. In sandwich immunoassays, less than 20% cross-reactivity with recombinant mouse IL-1ra is observed, less than 5% cross-reactivity with recombinant rat IL-1ra is observed, less than 2% cross-reactivity with recombinant porcine IL-1ra is observed, less than 0.4% cross-reactivity with recombinant human IL-1ra is observed, and less than 0.2% cross-reactivity with recombinant equine IL-1beta is observed.

Source

Polyclonal Goat IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant equine IL‑1ra/IL‑1F3
His26-Gln177
Accession # O18999

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.

Applications

Recommended Concentration

Sample

Western Blot

0.1 µg/mL

Recombinant Equine IL‑1ra/IL‑1F3 (Catalog # 2466-RA)

Immunocytochemistry

5-15 µg/mL

See below

Equine IL-1ra/IL-1F3 Sandwich Immunoassay

Recommended Concentration

Reagent

ELISA Capture (Matched Antibody Pair)

0.2-0.8 µg/mL

Use in combination with:

Detection Reagent: Equine IL‑1ra/IL‑1F3 Biotinylated Antibody (Catalog # BAF2466)

Standard: Recombinant Equine IL-1ra/IL-1F3 Protein (Catalog # 2466-RA)

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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IL‑1ra/IL‑1F3 in Equine PBMCs. IL-1ra/IL-1F3 was detected in immersion fixed equine peripheral blood mononuclear cells (PBMCs) using Goat Anti-Equine IL-1ra/IL-1F3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2466) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the Northern-Lights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to plasma membrane. 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

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: IL-1ra/IL-1F3

Secreted equine IL-1 receptor antagonist (IL-1ra) is a presumably 22‑25 kDa glycoprotein produced by variety of cell types that antagonizes IL-1 activity (1‑3). It is a member of the IL-1 family of proteins that includes IL-1 alpha and IL-1 beta. Although there is little amino acid (aa) identity (<30%) among the three IL-1 family members, all molecules bind to the same receptors, all show a beta -trefoil structure, and all are believed to have evolved from a common ancestral gene (1‑4). Equine IL-1ra is synthesized as a 177 aa precursor that contains a 25 aa signal sequence plus a 152 aa mature region. There is one intrachain disulfide bond and one potential N-linked glycosylation site (3, 5, 6). Mature equine sIL-1ra is 78%, 78%, 80%, 82%, and 76% aa identical to mature mouse, human, porcine, canine and bovine IL‑1ra, respectively. In human, three non-secreted IL-1ra isoforms have also been identified. It is unknown if such an analogous situation exists in equine. Cells known to secrete IL-1ra include fibroblasts, vascular smooth muscle cells, intestinal columnar epithelium, chondrocytes, macrophages, mast cells, neutrophils and hepatocytes.

There are two type I transmembrane glycoprotein receptors for IL-1ra. The first is the bioactive 80 kDa type I IL-1 receptor (IL-1 RI), and the second is the inert (decoy) 65 kDa type II IL-1 receptor. IL-1ra binding to IL-1 RI competitively blocks IL-1 ( alpha or beta ) binding to the same receptor. This results in receptor ligation without activation (1, 7). The type II IL-1 receptor is inert, and any binding of IL-1ra not only fails to block co-existing IL-1 activity, but may actually potentiate it by removing an IL-1 antagonist. Functionally, all activities attributed to IL-1ra are explained by its role as a competitive inhibitor of IL-1 binding to IL-1 RI (1, 2, 8, 9).

References