Equine IFN-gamma Antibody

Catalog #: AF1586 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF1586
AF1586-SP

IFN‑ gamma Inhibition of EMCV-induced Cytopathy and Neutralization by Equine IFN‑ gamma Antibody.

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Equine IFN-gamma Antibody Summary

Species Reactivity

Equine

Specificity

Detects equine IFN‑ gamma in ELISAs and Western blots. In sandwich immunoassays, approximately 20% cross‑reactivity with recombinant canine IFN‑ gamma is observed, less than 6% cross‑reactivity with recombinant bovine IFN‑ gamma and recombinant feline IFN‑ gamma is observed, and less than 0.2% cross‑reactivity with recombinant human IFN‑ gamma, recombinant mouse IFN‑ gamma, recombinant rat IFN‑ gamma, recombinant porcine IFN‑ gamma, recombinant rhesus macaque IFN‑ gamma, and recombinant cotton rat IFN‑ gamma is observed.

Source

Polyclonal Goat IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant equine IFN-gamma
Ala25-Gln166
Accession # P42160

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 Equine IFN-gamma (Catalog # 1586-HG)

Immunocytochemistry

5-15 µg/mL

See below

Equine IFN-gamma Sandwich Immunoassay

Recommended Concentration

Reagent

ELISA Capture (Matched Antibody Pair)

0.2-0.8 µg/mL

Use in combination with:

Detection Reagent: Equine IFN‑ gamma Biotinylated Antibody (Catalog # BAF1586)

Standard: Recombinant Equine IFN-gamma Protein (Catalog # 1586-HG)

Neutralization

Measured by its ability to neutralize IFN‑ gamma inhibition of EMCV-induced cytopathy in the L‑929 mouse fibroblast cell line. Vogel, S. and M. Hogan (1995) in Current Protocols in Immunology. Ciocio, R. (ed); John Wiley & Sons, Inc. p. 6. 9. 1. The Neutralization Dose (ND₅₀) is typically 0.25-1.25 µg/mL in the presence of 20 ng/mL Recombinant Equine IFN‑ gamma.

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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IFN‑ gamma Inhibition of EMCV-induced Cytopathy and Neutralization by Equine IFN‑ gamma Antibody. Recombinant Equine IFN-gamma (Catalog # 1586-HG) reduces the Encephalomyocarditis Virus (EMCV)-induced cytopathy in the L-929 mouse fibroblast cell line in a dose-dependent manner (orange line), as measured by Resazurin (Catalog # AR002). Inhibition of EMCV activity elicited by Recombinant Equine IFN-gamma (20 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Equine IFN-gamma Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1586). The ND₅₀ is typically 0.25-1.25 µg/mL.

Immunocytochemistry

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IFN‑ gamma in Equine PBMCs. IFN-gamma was detected in immersion fixed equine peripheral blood mononuclear cells (PBMCs) treated with Calcium Ionomycin and PMA using Goat Anti-Equine IFN-gamma Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1586) 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

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: IFN-gamma

Interferon-gamma (IFN-gamma ), also known as type II or immune interferon, exerts a wide range of immunoregulatory activities and is considered to be the prototype proinflammatory cytokine (1, 2). Mature equine IFN-gamma exists as a noncovalently linked homodimer of 20‑25 kDa variably glycosylated subunits (3, 4). It shares 73%‑82% amino acid sequence identity with bovine, canine, feline, and porcine IFN-gamma and 42%‑64% with cotton rat, human, mouse, rat, and rhesus IFN-gamma. IFN-gamma dimers bind to IFN-gamma RI (alpha subunits) which then interact with IFN-gamma RII (beta subunits) to form the functional receptor complex of two alpha and two beta subunits. Inclusion of IFN-gamma RII increases the binding affinity for ligand and the efficiency of signal transduction (5, 6). IFN-gamma is produced by a variety of immune cells under inflammatory conditions, notably by T cells and NK cells (7). It plays a key role in host defense by promoting the development and activation of Th1 cells, chemoattraction and activation of monocytes and macrophages, upregulation of antigen presentation molecules, and immunoglobulin class switching in B cells. It also exhibits antiviral, antiproliferative, and apoptotic effects (7, 8). In addition, IFN-gamma functions as an anti-inflammatory mediator by promoting the development of regulatory T cells and inhibiting Th17 cell differentiation (9, 10). The pleiotropic effects of IFN-gamma contribute to the development of multiple aspects of atherosclerosis (8).

References