Interferon gamma (IFNG) Mouse Monoclonal Antibody [Clone ID: #6]

PBS containing 50% glycerol, pH 7.2. No preservative is contained.

Store undiluted at 2-8°C for one month or (in aliquots) at -20°C for longer.
Avoid repeated freezing and thawing.

IFN-gamma, also known as type II interferon, is mainly produced in activated T lymphocytes and NK cells. It is involved in the regulation of nearly all phases of immune and inflammatory responses, such as the activation, growth, and differentiation of T cells, B cells, macrophages and also several other cell types, including endothelial cells and fibroblasts. IFN-gamma has antiviral and antiproliferative activities, it has also been shown to inhibit the proliferation of a variety of cells. Synthesis of IFN-gamma is upregulated by IL2, FGF basic and EGF.

Neutralization of IFN-gamma activity
1) Seed the FL cells in a 96 well microtiter plate at 1-2 x 10e4 cells/well and culture in MEM medium containing 5% FCS in microtiter plates for 2-3 days at 37oC in CO2 incubator.
2) Mix 30 IU/mL of IFN-gamma with same columes of 0, 5, 10 and 20 µg/mL of anti-human IFN-gamma antibodies in another microtiter plate and incubate the mixtures at 37oC for 1 hour.
3) Transfer the preincubated mixtures (100 µL/well) to the wells of FL cell, and incubate for over night.
4) Add sindbis virus (1 x 10e6 PFU/100 µL/well) suspension and incubate additionally for 22±2 hours at 37oC in CO2 incubator.
5) Aspirate the supernatant and add 0.02% Neutral Red solution to stain the viable cells after the virus challenge.
6) After the 40 minutes of incubation at 37oC in CO2 incubator, aspirate the dye, then add PBS (+) and incubate for another 40 minutes at room temperature.
7) Add 100 µL of 30% Ethanol containing 7 mM HCl to lyse the cells.
8) Measure the net cytopathic effect (CPE) by measuring the absorbance at 540 nm (A540).