Rhesus Macaque IFN-gamma Antibody

Catalog #: AF961 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF961
AF961-SP

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

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Rhesus Macaque IFN-gamma Antibody Summary

Species Reactivity

Rhesus Macaque

Specificity

Detects rhesus macaque IFN-gamma in ELISAs and Western blots. In sandwich immunoassays, less than 30% cross-reactivity with recombinant human IFN‑ gamma is observed and less than 0.1% cross-reactivity with recombinant mouse IFN‑ gamma, recombinant rat IFN‑ gamma, recombinant porcine IFN‑ gamma, recombinant canine IFN‑ gamma, recombinant cotton rat IFN‑ gamma, and recombinant feline IFN‑ gamma is observed.

Source

Polyclonal Goat IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant rhesus macaque IFN-gamma (R&D Systems, Catalog # 961-RM)
Gln24-Gln165
Accession # P63310

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.

Applications

Recommended Concentration

Sample

Western Blot

0.1 µg/mL

Recombinant Rhesus Macaque IFN-gamma (Catalog # 961-RM)

Rhesus Macaque IFN-gamma Sandwich Immunoassay

Recommended Concentration

Reagent

ELISA Capture (Matched Antibody Pair)

0.2-0.8 µg/mL

Use in combination with:

Detection Reagent: Rhesus Macaque IFN-gamma Biotinylated Antibody (Catalog # BAF961)

Standard: Recombinant Rhesus Macaque IFN-gamma Protein (Catalog # 961-RM)

Neutralization

Measured by its ability to neutralize IFN‑ gamma inhibition of EMCV-induced cytopathy in the HeLa human cervical epithelial carcinoma cell line. Meager, A. (1987) in Lymphokines and Interferons, a Practical Approach. Clemens, M.J. et al. (eds): IRL Press. 129. The Neutralization Dose (ND₅₀) is typically 1.5-5.0 µg/mL in the presence of 200 ng/mL Recombinant Rhesus Macaque 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 Primate IFN‑ gamma Antibody. Recombinant Rhesus Macaque IFN-gamma (Catalog # 961-RM) reduces the Encephalomyocarditis Virus (EMCV)-induced cytopathy in the HeLa human cervical epithelial carcinoma cell line in a dose-dependent manner (orange line), as measured by crystal violet staining. Inhibition of EMCV activity elicited by Recombinant Rhesus Macaque IFN-gamma (200 ng/mL) is neutralized (green line) by increasing concentrations of Rhesus Macaque IFN-gamma Antigen Affinity-purified Polyclonal Antibody (Catalog # AF961). The ND₅₀ is typically 1.5-5.0 µg/mL.

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 rhesus IFN-gamma exists as a noncovalently linked homodimer of 20-25 kDa variably glycosylated subunits (3). It shares 90% amino acid sequence identity with human IFN-gamma, 57-66% with bovine, canine, equine, feline, and porcine IFN-gamma, and 37-44% with cotton rat, mouse, and rat 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, antiproliferative, 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