All-in-one kit for robust detection of Phospho-STAT1 (Tyr701)
Phospho-STAT3 (Tyr705) cellular kit
This HTRF kit enables the cell-based quantitative detection of STAT3 phosphorylated on Tyr705, as a readout of the JAK/STAT signaling pathway.
- High sensitivity
- Faster and more convenient than ELISA
- Low sample consumption
HeLa cells were grown for 2 days in a T175 flask 37°C, 5%Co2. Stimulation was done with 5nM of IFNa for 15min. Cell culture medium was eliminated and cells supplemented with 3ml of lysis buffer before incubating for 45min. Soluble supernatants were collected after 10min centrifuging. Equal amounts of lysates were used for a side by side comparison of WB and HTRF. The two methods show a comparable sensitivity level: 750 cells. HTRF offers superior convenience over western blot.
HeLa cells (100,000 cells/well) were incubated for 1 hour at 37°C with various concentrations of antagonist. Agonist (IFNα) was then added and incubated for 30 minutes. After a 30-minute lysis incubation time, phosphorylated STAT3 was measured using the two-plate assay protocol of the HTRF phopsho-STAT3(Tyr705) cellular assay kit.
Hela cells (100,000 cells/well) were activated with IFNa for 15 min, using the two-plate assay protocol of the Phospho-STAT3 (Tyr705) and Total-STAT3 assays. Results obtained show a dose-response increase of STAT3 phosphorylation upon IFNα stimulation, while STAT3 expression level remains constant.
In response to cytokines and growth factors, STAT3 is phosphorylated by receptor-associated kinases and then forms dimers that translocate to the cell nucleus, where they act as transcription activators. STAT3 mediates the expression of a variety of genes in response to cell stimuli, and thus plays a key role in many cellular processes such as cell growth and apoptosis. The binding of Interleukin 6 - family cytokines to the gp130 receptor triggers STAT3 phosphorylation by JAK2. STAT3 is also a target of other receptors such as RTKs (EGFR, cmet.) and c-src.
Analyse of PI3K/AKT/mTor translational control pathway - Application Notes
Cell Signaling: Biomarkers, Phospho- & total-protein Assays - Flyers
physiologically relevant results fo fast flowing research - Flyers
Do all cell-based kinase assays perform similarly? - Posters
Simplify complex pathway dissection by combining the power of HTRF® cellular phospho-assays and the flexibility of the CyBi®-FeliX liquid handling system
PI3K/AKT/mTor translational control pathway - Posters
Analysis of a large panel of diverse biological samples and cellular models - Posters
Study a pathway of interest in PBMC and T cells - Application Notes
From 2D, 3D cell cultures to xenografts: A smart HTRF platform to maximize anticancer drug discovery
One technology across all samples - Application Notes
Tumor xenograft analysis: HTRF versus Western blot - Application Notes
Valuable guidelines for efficiently analyzing and interpreting results - Application Notes
Increased flexibility of phospho-assays - Application Notes
HTRF® phospho-protein platform facilitates the dissection of signaling pathways involved in insulin resistance and metabolic disorders
Multi-tissue cellular modeling and anlysis of insulin signaling - Posters
Insider Tips for successful sample treatment - Technical Notes
HTRF and WB compatible guidelines - Technical Notes
Mastering the art of cell signaling assays optimization - Guides
A solution for phospho-protein analysis in metabolic disorders - Posters
Detailed protocol and direct comparison with WB - Posters
Implement HTRF phospho-protein assays at every step of the drug discovery process, from in vitro to in vivo models
A single technology for 2D cells, 3D cells, and xenograft models - Posters
Protocol for tumor xenograft analysis with HTRF - Technical Notes
Unmatched ease of use, sensitivity and specificity assays - Videos
In collaboration with Bayer - Scientific Presentations
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