Human IFN gamma kit HTRF®

The HTRF human IFN gamma kit is designed for the quantification of human IFN gamma release in cell supernatant.

Voir plus d’informations
  • No-wash No-wash
  • Low sample consumption Low sample consumption
  • Under 2h bench time Under 2h bench time

The HTRF human IFN gamma kit is designed for the quantification of human IFN gamma release in cell supernatant.



IFN gamma is a pro-inflammatory cytokine secreted by T and NK cells in response to viral infection and other inflammatory conditions. IFN gamma plays a major role in innate and adaptive immunity by promoting T cell differentiation into TH1 and the development of Treg, as well as by inhibiting T cell differentiation into TH17. Its anti-tumor properties have been used in the treatment of osteoporosis and it is of great interest in immuno-oncology where it improves survival in patients with bladder or ovarian carcinoma.

Assessment of serum samples often requires enhanced sensitivity. In some cases, AlphaLISA assays may have sufficient sensitivity to enable the detection of low levels of analytes in serum or plasma. Check out the Human IFN-γ AlphaLISA kitand the Human IFN-γ biotin-free AlphaLISA kit’s analytical performances for more information or learn more about AlphaLISA. When assaying, always follow the recommended protocol and avoid highly haemolyzed samples.



Assay principle

Cell supernatant, sample, or standard is dispensed directly into the assay plate for the detection by HTRF® reagents (384-well low-volume white plate or Cisbio low-volume 96-well plate in 20 µl). The antibodies labeled with the HTRF donor and acceptor are pre-mixed and added in a single dispensing step, to further streamline the assay procedure. The assay can be run up to a 1536-well format by simply resizing each addition volume proportionally.

IFN gamma kit assay principle

Assay data analysis

The 4 Parameter Logistic (4PL) curve is commonly recommended for fitting an ELISA standard curve. This regression enables the accurate measurement of an unknown sample across a wider range of concentrations than linear analysis, making it ideally suited to the analysis of biological systems like cytokine releases.

To fully understand how to deal with HTRF data processing and also 4PL 1/y² fitting, please visit this page.

Cisbio also worked with Myassays.com to help you in your data analysis. By clicking on this link, you’ll be able to access a free online software to run your IFN gamma analysis.

Technical specifications of human IFN gamma kit

Sample size16 µL
Final assay volume20 µL
Kit componentsLyophilized standard, frozen detection antibodies, buffers &protocol.
LOD &LOQ (in Diluent)14 pg/mL &21 pg/mL
Range21 – 4,000 pg/mL
Time to resultOvernight at RT
CalibrationNIBSC (82/587) value (IU/mL) = 0,019 x HTRF hIFN? value (pg/mL)
SpeciesHuman only

Intra and inter assay

Intra-assay (n=24)

SampleMean [IFN?] (pg/mL)CV

Mean CV9%

Each of the 3 samples was measured 24 times, and % CV was calculated for each sample.

Inter-assay (n=4)

Sample[IFNg] (pg/mL)Mean (delta R)CV

Mean CV9.7%

Each of the samples was measured in 4 different experiments, and % CV was calculated for each sample.

Dilutional linearity

​The excellent % of recovery obtained from these experiments show the good linearity of the assay.​

Dilution factor[IFNg] expected (pg/mL)[IFNg] detected (pg/mL)Recovery

Spike and recovery

Sample[IFNg] added (pg/mL)[IFNg] expected (pg/mL)[IFNg] detected (pg/mL)Recovery

The same amount of recombinant cytokine was added to 2 different serum samples, and the set of responses obtained from a standard curve was compared to the calculated expected values. The ~ 100% of recovery observed validates the sample matrix used for this assay.

IFNg secretion in PBMCs stimulated with PMA and Ionomycin

PBMC plated at 50, 100, 200 and 400 kcells/well were stimulated for 3h with increasing concentrations of PMA (0, 1, 50 ng/mL) added to a 500 ng/mL Ionomycin solution. 16 µL of supernatants were then transferred into a white detection plate (384, low volume) to be analyzed with the Human IFN? Assay.

IFN? secretion in PBMCs stimulated with PMA and Ionomycin

Following T-cell activation and differenciation with HTRF reagents: IL-2, IFN-g and IL-17

In collaboration with Novartis - Présentations scientifiaues

HTRF Cytokine cell-based assays

Monitor immune regulation the simple way - Flyers

Inflammation cell by cell

HTRF solutions for each cell type - Flyers

HTRF assays for Oncology and Inflammation

Signaling in the immune system - Brochures

HTRF measurement of cytokine release from fresh blood samples

In collaboration with Blood assay solution - Notes d'application

Cytokine secretion profile on PBMCs

One sample to measure them all - Posters

Guidelines from PBMC isolation to cytokine assay optimisation

Perform and optimize cytokine assays on PBMC - Notes techniques

How to freeze PBMC

Validated protocol - Notes techniques

How to measure multiple cytokines from 1 sample

What to expect at the bench - Vidéos

HTRF makes cytokine detection easy

A fun video introducing you to cytokines assays with HTRF - Vidéos

STING HTRF offer to bridge innate and adaptive immunity

cGAS-STING signaling pathway from A to Z - Brochures

Cisbio Product Catalog 2019

All your HTRF assays in one document! - Catalogue

A guide to Homogeneous Time Resolved Fluorescence

General principles of HTRF - Guides

Functional response to PD1 blockade rapidly assessed through HTRF IFNγ quantification

For researcher working on checkpoint inhibitors - Notes d'application

Cytokine assays: a guide to success with HTRF

Setting up a successful cytokine assay - Guides

Novel HTRF platform to delineate STING pathway

Explore the whole STING pathway with a single technology - Posters

Webinar: What's next for CAR T-cells immunotherapy

Featuring a panel of experts - Vidéos

Portfolio of HTRF cytokine assays

This leaflet presents the analytical performances of each assay - Brochures

Cytokine Portfolio Flyer

Discover no-wash cytokine assay solutions - Flyers

Innovative solutions in Virology research

Kits and reagents for virology research - Flyers

Homogeneous PPi assays for virology research

A review compiling published scientific data about virology research - Notes d'application

Assays for autoimmunity research

Advance your research on autoimmune diseases - Flyers

Review of cell-based methods to monitor CAR-T activation and cell killing effectiveness

A novel CAR-T cell therapy based on chimeric antigen receptors for tumor cells, that target CD19 antigens. - Notes d'application

Immuno-Oncology Drug Discovery

Explore our solutions for Immuno-oncology research - Flyers

Safety Data Sheet IFN g (h) Kit / 62HIFNGPEG

62HIFNGPEG - Fiches de sécurité

Plate Reader Requirement

Choosing the right microplate reader ensures you’ll get an optimal readout. Discover our high performance reader, or verify if your lab equipment is going to be compatible with this detection technology.

Let's find your reader