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Mouse IFN beta kit HTRF®

The HTRF mouse IFN beta kit is designed for the quantification of mouse IFN beta release in cell supernatant.

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  • No-wash No-wash
  • Low sample consumption Low sample consumption
  • Under 2h bench time Under 2h bench time

The HTRF mouse IFN beta kit is designed for the quantification of mouse IFN beta release in cell supernatant.

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In stock

Overview

Type I interferons, interferon alpha (IFN-α) and interferon beta (IFN-β), play essential roles in the innate immune response to viral infections. IFN-β is produced by fibroblasts, macrophages, and dendritic cells, as well as endothelial cells. The expression of the IFN-β gene is controlled by transcription factors like IRF-3, c-Jun/ATF-2, and NF-kappaB downstream of Pattern Recognition Receptors, for example Toll-like receptors. In particular, IFN-β is the primary functional readout of cGas-cGAMP-STING pathway activation. Once secreted, IFN-β binds and activates the IFNAR1/IFNAR2 receptor complex, which relies on STAT1/2 transcription factors. Phosphorylated STAT1/2 leads to the induction of anti-viral gene expression.

Benefits

  • SCALABLE FROM 96 TO 1536 WELLS
  • PANEL QUANTIFICATION

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.

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

Technical specifications of mouse IFN beta kit

Sample size16 µL
Final assay volume20 µL
Kit componentsLyophilized standard, frozen detection antibodies, buffers, & protocol
LOD & LOQ (in Diluent)4 pg/mL & 17 pg/mL
Range 17 - 2,000 pg/mL
Time to result Overnight at RT
SpeciesMouse only

Intra and inter assay

Intra assay (n=24)

Sample

Mean [IFN-β] (pg/mL)

CV

1

75

10%

2

200

3%

3

1125

3%


Mean CV

5.3%

Inter assay (n=6)

Sample

Mean [IFN-β] (pg/mL)

CV

1

58

3.6%

2

348

3.3%

3

1092

7.3%


Mean CV

4.7%

Spike and recovery

A fixed concentration of recombinant mouse IFN-beta was spiked into several cell supernatants containing known concentrations of mouse IFN-beta. Samples were mixed at a 1:1 volume ratio. The set of responses obtained from the standard curve was compared to the calculated expected values.

Dilutional linearity

A cell supernatant containing a known concentration of mouse IFN-beta was serially diluted in the culture media used to generate the sample. The set of responses obtained from the mouse IFN-beta standard curve was compared to the calculated expected values.

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The mouse STING-specific agonists CMA and DMXAA trigger IFN-β release from NIH-3T3 fibroblast cells

The mouse NIH-3T3 fibroblast cell line was plated in a 96-well culture plate (100,000 cells/well) and incubated for 6h. The cells were then treated with 200 µM of the synthetic mouse STING-specific agonists CMA and DMXAA (under 100 µL). After 24h incubation, cell supernatants were collected and secreted levels of IFN-beta were measured following the procedure given in the HTRF kit’s package insert.

Mouse IFN-beta kit_NIH-3T3 cells treated with CMA and DMXAA

IFN-β secretion by the murine RAW 264.7 macrophage cell line treated with several STING agonists

The murine RAW 264.7 macrophage cell line was plated in a 96-well culture plate (100,000 cells/well) and incubated overnight. The cells were then treated with a fixed concentration of the synthetic STING agonists CMA, DMXAA, and ADUS100, as well as with the endogenous STING agonist 2'3'-cGAMP (under 100 µL). After 24h secretion, cell supernatants were collected and levels of released IFN-beta were measured following the procedure given in the HTRF kit’s package insert.

HTRF mouse IFN-beta kit_RAW 264.7 cells treated with several STING agonists

Dose-dependent secretion of IFN-β by the murine RAW 264.7 macrophage cell line treated with 2’3’-cGAMP and ADUS100

The murine RAW 264.7 macrophage cell line was plated in a 96-well culture plate (100,000 cells/well) and incubated for 5h. The cells were then treated with increasing concentrations of either the endogenous STING agonist 2'3'-cGAMP, or the synthetic small molecule ADUS100 (under 100 µL). After an overnight secretion, cell supernatants were collected and levels of released IFN-beta were measured following the procedure given in the HTRF kit’s package insert.​

HTRF mouse IFN-beta kit_RAW 264.7 cells treated with 2’3’-cGAMP and ADUS100

Product Insert IFN b (h) Kit / 62HIFNBPEG-62HIFNBPEH

62HIFNBPEG-62HIFNBPEH - Product Insert

How to measure multiple cytokines from 1 sample

What to expect at the bench - Videos

How to freeze PBMC

Guideline for PBMC freezing - Technical Notes

Guidelines from PBMC isolation to cytokine assay optimisation

Perform and optimize cytokine assays on PBMC - Technical Notes

Cytokine secretion profile on PBMCs

One sample to measure them all - Posters

HTRF measurement of cytokine release from fresh blood samples

In collaboration with Blood assay solution - Application Notes

Normalize HTRF cytokine assays with viability assay on Spectramax I3X-Minimax

In collaboration with Molecular Devices - Application Notes

HTRF assays for Oncology and Inflammation

Signaling in the immune system - Brochures

HTRF Cytokine cell-based assays

Monitor immune regulation the simple way - Flyers

HTRF makes cytokine detection easy

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

STING HTRF offer to bridge innate and adaptive immunity

cGAS-STING signaling pathway from A to Z - Brochures

Novel HTRF platform to delineate STING pathway

Explore the whole STING pathway with a single technology - Posters

HTRF Product Catalog 2020 July update

All your HTRF assays in one document! - Catalog

A guide to Homogeneous Time Resolved Fluorescence

General principles of HTRF - Guides

Cytokine assays: a guide to success with HTRF

Setting up a successful cytokine assay - Guides

Portfolio of HTRF cytokine assays

This leaflet presents the analytical performances of each assay - Brochures

How HTRF compares to Western Blot and ELISA

Get the brochure about technology comparison. - Brochures

Characterize STING agonists with HTRF human IFNb

Data about HTRF IFNb correlation with gene reporter assay and ELISA - Application Notes

Note about HMGB1 assay

HTRF HMGB1, IL-1β and IL-18 assays discriminate between inflammasome triggered pyroptosis, and necroptosis - Application Notes

Homogeneous PPi assays for virology research

A review compiling published scientific data about virology research - Application Notes

Product Insert IFN b (m) Kit / 62MIFNBPEG-62MIFNBPEH

62MIFNBPEG-62MIFNBPEH - Product Insert

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