Phospho-STAT3 (Tyr705) cellular kit
All-in-one kit for robust detection of Phospho-STAT3 (Ser423/425)
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The HTRF mouse IFN beta kit is designed for the quantification of mouse IFN beta release in cell supernatant.
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.
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.
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.
Sample size | 16 µL |
Final assay volume | 20 µL |
Kit components | Lyophilized 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 |
Species | Mouse only |
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% |
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.
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.
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.
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.
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.
How to measure multiple cytokines from 1 sample
What to expect at the bench - Videos
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
HTRF assays for Oncology and Inflammation
Signaling in the Immune System - Brochures
HTRF Cytokine cell-based assays
Monitor immune regulation the simple way - Flyers
In collaboration with CNRS - Scientific Presentations
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
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 Life Sciences 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
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
Assays for autoimmunity research
Advance your research on autoimmune diseases - Flyers
Advance your research on Fibrosis
Kits and reagents for Fibrosis research - Flyers
Safety Data Sheet (DEU) IFN b (m) Kit / 62MIFNBPEG
62MIFNBPEG - Safety Data Sheet
Safety Data Sheet (ELL) IFN b (m) Kit / 62MIFNBPEG
62MIFNBPEG - Safety Data Sheet
Safety Data Sheet (FRA-FR) IFN b (m) Kit / 62MIFNBPEG
62MIFNBPEG - Safety Data Sheet
Safety Data Sheet (ITA) IFN b (m) Kit / 62MIFNBPEG
62MIFNBPEG - Safety Data Sheet
Safety Data Sheet (SPA) IFN b (m) Kit / 62MIFNBPEG
62MIFNBPEG - Safety Data Sheet
Safety Data Sheet (ENG-GB) IFN b (m) Kit / 62MIFNBPEG
62MIFNBPEG - Safety Data Sheet
Safety Data Sheet (ENG-US) IFN b (m) Kit / 62MIFNBPEG
62MIFNBPEG - Safety Data Sheet
Safety Data Sheet (DEU) IFN b (m) Kit / 62MIFNBPEH
62MIFNBPEH - Safety Data Sheet
Safety Data Sheet (ELL) IFN b (m) Kit / 62MIFNBPEH
62MIFNBPEH - Safety Data Sheet
Safety Data Sheet (FRA-FR) IFN b (m) Kit / 62MIFNBPEH
62MIFNBPEH - Safety Data Sheet
Safety Data Sheet (ITA) IFN b (m) Kit / 62MIFNBPEH
62MIFNBPEH - Safety Data Sheet
Safety Data Sheet (SPA) IFN b (m) Kit / 62MIFNBPEH
62MIFNBPEH - Safety Data Sheet
Safety Data Sheet (ENG-GB) IFN b (m) Kit / 62MIFNBPEH
62MIFNBPEH - Safety Data Sheet
Safety Data Sheet (ENG-US) IFN b (m) Kit / 62MIFNBPEH
62MIFNBPEH - Safety Data Sheet
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