IFN-γ ELISA Kits

Interferon-γ (IFN-γ) is a cytokine classified as a type II interferon. It was discovered as an antiviral factor produced by leukocytes stimulated with phytohemagglutinin, a type of mitogen¹⁾. Human IFN-γ consists of 143 amino acids, while mouse IFN-γ consists of 133 amino acids; both function as homodimers. The IFN-γ receptor is composed of the IFNGR1 and IFNGR2 subunits; binding of IFN-γ activates the downstream JAK–STAT signaling pathway. Activated STAT1 forms a homodimer, translocates into the nucleus, and binds to a DNA sequence known as the IFN-γ activated site (GAS), thereby inducing downstream gene expression.
IFN-γ is an important cytokine involved in both innate and adaptive immunity. Immune cells secrete IFN-γ in response to cytokines (such as IL-12), pathogens, or tumor-derived substances. Cells known to produce IFN-γ include natural killer (NK) cells and helper T (Th1) cells. Secreted IFN-γ induces a wide range of immune responses, including antiviral activity, activation of macrophages, maturation of dendritic cells, and induction of T-cell differentiation²⁾. IFN-γ also plays a significant role in antitumor immunity, with reported effects including suppression of metastasis, induction of apoptosis, and inhibition of angiogenesis²⁾.

LBIS™ Human IFN-γ ELISA Kit is a sandwich ELISA kit for the quantitative measurement of human IFN-γ. This kit enables highly sensitive quantification of IFN-γ in biological samples. The standard included in the kit is a recombinant protein produced in Escherichia coli.

Kit Performance

Assay target	IFN-γ
Calibration curve range	0.768-75.0 pg/mL
Analysis sample	Human Serum/Plasma (EDTA・Heparin) /Cell culture supernatant*
Sample volume	50 μL (2x dilution)
Measurement duration	Approx. 2 hours and 20 minutes
Wavelength	Primary wavelength : 450 nm Reference wavelength : 620 nm

*Measurement availability depends on the culture medium.
When using cell culture supernatant as a sample, please make sure to perform recovery test and dilution linearity test on the culture medium before use.

Example of Calibration Curve

Measurement of IFN-γ in Cell Culture Supernatants of Human Peripheral Blood Mononuclear Cells (PBMCs)

IFN-γ in cell culture supernatants of human PBMCs was measured using this product and a comparable kit from another manufacturer.

＜Experimental conditions＞

Control :

RPMI-1640 + 10% FBS

Without PHA stimulation :

Culture supernatant of PBMCs (5x10⁵ cells/mL) cultured for 3 days in RPMI-1640 + 10% FBS

With PHA stimulation :

Culture supernatant of PBMCs (5x10⁵ cells/mL) cultured for 3 days in RPMI-1640 + 10% FBS
Stimulated with 10 µg/mL phytohemagglutinin (PHA) during culture

[Result]
Using this product and the kit from another manufacturer, an increase in IFN-γ in human PBMC culture supernatants upon stimulation with 10 µg/mL PHA was confirmed.

LBIS™ Mouse IFN-γ ELISA Kit is a sandwich ELISA kit for the quantitative measurement of mouse IFN-γ. This kit enables highly sensitive quantification of IFN-γ in biological samples. The standard included in the kit is a recombinant protein produced in Escherichia coli.

Kit Performance

Assay target	IFN-γ
Calibration curve range	2.05-500 pg/mL
Analysis sample	Mouse Serum/Plasma (EDTA・Heparin)
Sample volume	25 μL (2x dilution)
Measurement duration	Approx. 3 hours and 50 minutes
Wavelength	Primary wavelength : 450 nm Reference wavelength : 620 nm

Example of Calibration Curve

Measurement of IFN-γ in Mouse Serum and Plasma

Sample: BALB/c, male, 8-weeks old, 16-hour fast

[Result]
This product was able to detect IFN-γ even in samples that were below the lower limit of quantification of another manufacturer's kit.

1. Wheelock, E. F.: Science, 149(3681), 310(1965).
   Interferon-like virus-inhibitor induced in human leukocytes by phytohemagglutinin
2. Jorgovanovic, D. et al.: Biomark. Res., 8, 49(2020).
   Roles of IFN-γ in tumor progression and regression: a review