Porcine IFN-gamma Antibody

Catalog #: AF985 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF985
AF985-SP

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

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Porcine IFN-gamma Antibody Summary

Species Reactivity

Porcine

Specificity

Detects porcine IFN‑ gamma in direct ELISAs and Western blots.

Source

Polyclonal Goat IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant porcine IFN-gamma
Ser21-Lys166
Accession # P17803

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 Porcine IFN-gamma (Catalog # 985-PI)

ELISA

This antibody functions as an ELISA detection antibody when paired with Mouse Anti-Porcine IFN‑ gamma Monoclonal Antibody(Catalog # MAB9852).

This product is intended for assay development on various assay platforms requiring antibody pairs. We recommend the Porcine IFN-gamma DuoSet ELISA Kit (Catalog # DY985) for convenient development of a sandwich ELISA.

Neutralization

Measured by its ability to neutralize IFN‑ gamma inhibition of EMCV-induced cytopathy in the PK‑15 porcine kidney epithelial cell line. The Neutralization Dose (ND₅₀) is typically 0.6-3.0 µg/mL in the presence of 0.2 ng/mL Recombinant Porcine 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 Porcine IFN‑ gamma Antibody. Recombinant Porcine IFN-gamma (985-PI) reduces the Encephalomyocarditis Virus (EMCV)-induced cytopathy in the PK-15 porcine kidney epithelial cell line in a dose-dependent manner (orange line), as measured by Resazurin (AR002). Inhibition of EMCV activity elicited by Recombinant Porcine IFN-gamma (0.2 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Porcine IFN-gamma Antigen Affinity-purified Polyclonal Antibody (Catalog # AF985). The ND₅₀ is typically 0.6-3.0 µg/mL.

ELISA

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Porcine IFN‑ gamma ELISA Standard Curve. Recombinant Porcine IFN-gamma protein was serially diluted 2-fold and captured by Mouse Anti-Porcine IFN-gamma Monoclonal Antibody (MAB9852) coated on a Clear Polystyrene Microplate (DY990). Goat Anti-Porcine IFN-gamma Antigen Affinity-purified Polyclonal Antibody (Catalog # AF985) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (DY998) followed by Substrate Solution (DY999) and stopping the enzymatic reaction with Stop Solution (DY994).

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 porcine IFN-gamma exists as a noncovalently linked homodimer of 20‑25 kDa variably glycosylated subunits (3). It shares 72%‑79% amino acid sequence identity with bovine, canine, equine, and feline IFN-gamma and 41%‑57% 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 (4, 5). IFN-gamma is produced by a variety of immune cells under inflammatory conditions, notably by T cells and NK cells (6). 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, anti-proliferative, and apoptotic effects (6, 7). In addition, IFN-gamma functions as an anti-inflammatory mediator by promoting the development of regulatory T cells and inhibiting Th17 cell differentiation (8, 9). The pleiotropic effects of IFN-gamma contribute to the development of multiple aspects of atherosclerosis (7).

References