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Human & Mouse IFNγ assays

HTRF® human & mouse IFNγ kits are rapid sandwich assays for sensitive and accurate interferon-γ (IFNγ) quantification on cell culture supernatants or whole cells. The simple and robust procedure gives increased throughput compared to ELISA. IFNy production can easily be monitored in a high throughput cell-based screening.

 

Cytokines are of particular interest in the fields of oncology, infectious diseases, allergies and autoimmune diseases. Cisbio Bioassays has developed a line of reagents for quantifying a variety of cytokines, based on our patented HTRF® technology.

Features

  • Cost effective alternative to ELISA
  • Easy to use: one addition, no washing steps
  • Highly sensitive
  • Small sample size: down to 5µL
  • Easy miniaturization & automation
  • Robust: compatible with cell supernatant and whole cells

Applications

  • IFNγ quantification in human and mouse cell culture or whole cells
  • Compound screening using IFNγ secreting cells
  • HTS for inhibitors of IFNγ production on NK cells and Th1 cells ((Enomoto et al, 2000)

  • Allergy, Inflammation

 

Human and mouse IFNγ kits are based on the same assay principle. Both assays use a sandwich immunoassay involving two specific antibodies, one anti-IFNγ labelled with Eu-Cryptate and the second anti-IFNγ labelled with XL665 or d2. The signal is proportional to the concentration of the antigen present in the sample.

 

Human and mouse IFNγ tests, like all cytokine assays, can be run with cell supernatants or whole cells using the protocols described here. The antibodies labelled with HTRF® fluorophores may be pre-mixed and added in a single dispensing step to further streamline the protocol. The assay can be run in 96- to 1536-well plates by simply resizing each addition volume proportionally. The mouse IFNγ assay protocol is described here, using a white 384-well small volume plate.

Human IFN y standard curve

  Human IFNγ Mouse IFNγ
Incubation 18 - 24h at RT 2h at RT
Standard range 23.4 to 1,500 pg/mL 7.8 to 2,000 pg/mL
Detection limit 10 pg/mL
NIBSC 88/606		 30 pg/mL
Linear range

Up to 5,000 pg/mL

 Up to 2,000 pg/mL

The standard curves for each assay were run in white 384-well small volume plates and read on PHERAstarFS Laser.

 

Mouse IFN y standard curve

Cisbio Bioassays' assays are ideal for highly sensitive cytokine screening. They have rapid, easily miniaturized protocols that can be run in a single plate from cell stimulation through to assay measurement. Figure 1 shows levels of human TNFα (panel A), human IFNγ (panel B), human IL1β (panel C) and human IL6 (panel D) detected using the cell-based and supernatant assay formats. These data show that both protocols give similar results and consistent cytokine concentrations with respect to the number of cells per well, indicating that the presence of cells does not alter assay performance. Levels of IL6 (panel D) were not quantified above 6,250 cells per well, as all concentrations were off the scale.

Cytokine performance

Ordering Info

DescriptionCat. noProduct insertMSDS
Human IFN gamma kit - 1,000 tests62IFNPEBpdfpdf
Human IFN gamma kit - 20,000 tests62IFNPECpdfpdf
Mouse IFN gamma kit - 1,000 tests6FMIFPEB-pdf
Mouse IFN gamma kit - 10,000 tests6FMIFPEH-pdf

Companion products

DescriptionCat. noProduct insertMSDS
Diluent 1 - 20 ml62DL1DDD-pdf
Human IFN gamma calibrator62IFNCDA-pdf
Human IFN gamma control62IFNTDA-pdf
Reconstitution buffer 2 - 13 ml62RB2RDD--

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