应用

建议稀释比

免疫印迹 (WB)

-

免疫细胞化学 (ICC/IF)

-

抑制试剂 (IA)

-

酶联免疫吸附实验 (ELISA)

5 µg/mL

中和实验 (Neu)

Assay-Dependent

体外检测 (IV)

-

功能检测 (Functional)

Assay-Dependent

流式细胞分析 (Flow)

-

产品详细信息

Description: The NIB42 antibody reacts with human interferon-gamma (IFN-gamma). The NIB42 antibody is a neutralizing antibody. IFN-gamma is an ~20 kDa factor produced by activated T, B and NK cells and is an anti-viral and anti-parasitic cytokine. IFN-gamma in synergy with other cytokines, such as TNF-alpha, inhibits proliferation of normal and transformed cells. Immunomodulatory effects of IFN-gamma are exerted on a wide range of cell types expressing the high affinity receptors for IFN-gamma. Glycosylation of IFN-gamma does not affect its biological activity.

Applications Reported:The NIB42 antibody has been reported for use in neutralization of human IFN gamma and for ELISA capture.

Applications Tested: The Functional Grade Purified NIB42 antibody has been tested by LAL assay to verify low endotoxin levels and has been tested for ELISA capture and in bioassay for neutralization of IFN gamma bioactivity. The NIB42 antibody at 5 µg/mL has been found to inhibit by 50% the biological effects of 1 ng/mL human IFN gamma(ND50), in an EMCV assay of A549 cell protection. Detailed information and protocols about cytokine bioassays and in vitro cytokine neutralization using antibodies can be found in the BestProtocols® section.

The NIB42 antibody has been tested as the capture antibody in a sandwich ELISA for analysis of human Interferon-gamma (IFN-g) in combination with the biotin 4S.B3 (13-7319) antibody for detection and recombinant human IFNg (39-8319) as the standard. A suitable range of concentrations of this antibody for ELISA capture is 2-8 µg/mL. A standard curve consisting of doubling dilutions of the recombinant standard over the range of 1000 pg/mL - 8 pg/mL should be included in each ELISA plate. For ELISPOT capture, the alternative clone MD-1 is recommended.

Storage and handling: Use in a sterile environment.

Filtration: 0.2 µm post-manufacturing filtered.

Purity: Greater than 90%, as determined by SDS-PAGE.

Endotoxin Level:Less than 0.001 ng/µg antibody, as determined by LAL assay.

Aggregation:Less than 10%, as determined by HPLC.

靶标信息

IFN gamma (Interferon gamma, Type II interferon) is a macrophage activation factor, and immune interferon that is produced primarily by T-lymphocytes and natural killer cells in response to antigens, mitogens, Staphylococcus enterotoxin B, phytohemaglutanin and other cytokines. IFN gamma is a dimeric protein consisting of two 146 amino acid subunits. IFN gamma is a glycoprotein that exists, functionally, as a homodimer of approximately 45 kDa. On SDS-PAGE, IFN gamma appears as a combination of 25, 20 and minor 15.5 kDa bands as a result of differential glycosylation. The biological activity of the IFN gamma homodimer is highly species specific. Human IFN gamma does not show cross-reactivity with mouse. IFN gamma function includes the following: antiviral activity, tumor antiproliferative activity, induction of class I and II MHC, macrophage activation, and enhanced immunoglobulin secretion by B lymphocytes. IFN gamma is involved in cytokine regulation and also acts synergistically with other cytokines. Activation of IFN gamma takes place through binding of IFN gamma receptor I and II, and activating the JAK-STAT pathway. IFN gamma does not show any homology with IFN alpha or IFN beta but human IFN gamma shows about 40% sequence homology with mouse IFN gamma. IFN gamma is upregulated by IL2, FGF basic, EGF and downregulated by vitamin D3 or DMN. IFN gamma gene mutations are associated with aplastic anemia.