Phospho-STAT1 (Tyr701) cellular kit
All-in-one kit for robust detection of Phospho-STAT1 (Tyr701)
Phospho-STAT3 (Tyr705) cellular kit HTRF®
This HTRF kit enables the cell-based quantitative detection of STAT3 phosphorylated on Tyr705, as a readout of the JAK/STAT signaling pathway.
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Ready-to-use
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High sensitivity
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Faster and more convenient than ELISA
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Low sample consumption
Overview
This HTRF cell-based assay enables rapid, quantitative detection of STAT3 phosphorylated on Tyr705 as a readout of JAK/STAT signaling activity. STAT3 is upregulatd in many cancers, making it a powerful tool in anti-cancer research.
Benefits
- SPECIFICITY
- PRECISION
Phospho-STAT3 (Tyr705) assay principle
The Phospho-STAT3 (Tyr705) assay measures STAT3 when phosphorylated at Tyr705. Contrary to Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis or transfer. The Phospho-STAT3 (Tyr705) assay uses 2 labeled antibodies: one with a donor fluorophore, the other one with an acceptor. The first antibody is selected for its specific binding to the phosphorylated motif on the protein, the second for its ability to recognize the protein independent of its phosphorylation state. Protein phosphorylation enables an immune-complex formation involving both labeled antibodies and which brings the donor fluorophore into close proximity to the acceptor, thereby generating a FRET signal. Its intensity is directly proportional to the concentration of phosphorylated protein present in the sample, and provides a means of assessing the proteins phosphorylation state under a no-wash assay format.
Phospho-STAT3 (Tyr705) 2-plate assay protocol
The 2 plate protocol involves culturing cells in a 96-well plate before lysis then transferring lysates to a 384-well low volume detection plate before adding phospho-STAT3 (Tyr705) HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.
Phospho-STAT3 (Tyr705) 1-plate assay protocol
Detection of Phosphorylated STAT3 (Tyr705) with HTRF reagents can be performed in a single plate used for culturing, stimulation and lysis. No washing steps are required. This HTS designed protocol enables miniaturization while maintaining robust HTRF quality.
HTRF compared to Western Blot using phospho-STAT3 assay
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.
IL6 dose-response on NIH 3T3 cells
Murine NIH 3T3 cells (100,000 cells/well) were stimulated for 30 minutes at 37°C with various concentrations of IL6. 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.
Inhibition effect of JAK inhibitor on HeLa cells
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.
HTRF total-STAT3 assay used to check the phosphorylation status of STAT3
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.
STAT3 in cytokine receptor signaling
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.
Simplified pathway dissection with HTRF phospho-assays and CyBi-felix liquid handling
Analyse of PI3K/AKT/mTor translational control pathway - Application Notes
Investigating kinase activity in a cellular context
HTRF cellular assays - Scientific Presentations
Open R&D: Sanofi Access Platform
In collaboration with Sanofi - Scientific Presentations
Cell Signaling: Biomarkers, Phospho- & total-protein Assays - Flyers
HTRF cellular phospho-protein assays
Physiologically relevant results fo fast flowing research - Flyers
Biomarker and Cell Signaling Assays for Fibrosis and NASH
HTRF solutions for NASH - Flyers
HTRF solutions for each cell type - Flyers
Cell Signaling: Biomarkers, Phospho- & total-protein assays - Flyers
PI3K/AKT/mTor translational control pathway - Posters
Universal HTRF® phospho-protein platform: from 2D, 3D, primary cells to patient derived tumor cells
Analysis of a large panel of diverse biological samples and cellular models - Posters
TCR signaling investigation with HTRF phospho assays
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
HTRF phospho assays reveal subtle drug induced effects in tumor-xenografts
Tumor xenograft analysis: HTRF versus Western blot - Application Notes
HTRF cell-based phospho-protein data normalization
Valuable guidelines for efficiently analyzing and interpreting results - Application Notes
HTRF phospho-total lysis buffer: a universal alternative to RIPA lysis buffers
Increased flexibility of phospho-assays - Application Notes
Multi-tissue cellular modeling and anlysis of insulin signaling - Posters
Best practices for analyzing brain samples with HTRF® phospho assays for neurosciences
Insider Tips for successful sample treatment - Technical Notes
HTRF Alpha-tubulin Housekeeping kit
Properly interpret your compound effect - Application Notes
Optimize your HTRF cell signaling assays on tissues
HTRF and WB compatible guidelines - Technical Notes
Key guidelines to successful cell signaling experiments
Mastering the art of cell signaling assays optimization - Guides
HTRF® cell signaling platform combined with iCell® Hepatocytes
A solution for phospho-protein analysis in metabolic disorders - Posters
HTRF phospho-assays reveal subtle drug-induced effects
Detailed protocol and direct comparison with WB - Posters
A single technology for 2D cells, 3D cells, and xenograft models - Posters
Best practices for analyzing tumor xenografts with HTRF phospho assays
Protocol for tumor xenograft analysis with HTRF - Technical Notes
How to run a cell based phospho HTRF assay
What to expect at the bench - Videos
Unleash the potential of your phosphorylation research with HTRF
Unmatched ease of use, sensitivity and specificity assays - Videos
Cell-based kinase assays in HTS ? potential and limitations for primary and secondary screening
In collaboration with Bayer - Scientific Presentations
Product Insert STAT3 P-Y705 Kit / 62AT3PET
62AT3PET - Product Insert
Product Insert STAT3 P-Y705 Kit / 62AT3PEG-62AT3PEH
62AT3PEG-62AT3PEH - Product Insert
All your HTRF assays in one document! - Catalog
A guide to Homogeneous Time Resolved Fluorescence
General principles of HTRF - Guides
How HTRF compares to Western Blot and ELISA
Get the brochure about technology comparison. - Brochures
Unleash the potential of your phosphorylation research with HTRF
A fun video introducing you to phosphorylation assays with HTRF - Videos
How to run a cell based phospho HTRF assay
3' video to set up your Phospho assay - Videos
Innovative solutions in Virology research
Kits and reagents for virology research - Flyers
Guidelines for Cell Culture and Lysis in Different Formats Prior to HTRF Detection
Seeding and lysing recommendations for a number of cell culture vessels. - Technical Notes
Assessment of drug efficacy and toxicity by combining innovative technologies
Combination of AlphaLISA®, HTRF®, or AlphaLISA® SureFire® Ultra™ immunoassays with the ATPlite™ 1step cell viability assay - Application Notes
Methodological Aspects of Homogeneous Time-Resolved Fluorescence (HTRF)
Learn how to reduce time and sample consumption - Application Notes
Safety Data Sheet STAT3 P-Y705 Kit / 62AT3PEG
62AT3PEG - Safety Data Sheet
Safety Data Sheet (DEU) STAT3 P-Y705 Kit / 62AT3PEG
62AT3PEG - Safety Data Sheet
Safety Data Sheet (ELL) STAT3 P-Y705 Kit / 62AT3PEG
62AT3PEG - Safety Data Sheet
Safety Data Sheet (FRA-FR) STAT3 P-Y705 Kit / 62AT3PEG
62AT3PEG - Safety Data Sheet
Safety Data Sheet (ITA) STAT3 P-Y705 Kit / 62AT3PEG
62AT3PEG - Safety Data Sheet
Safety Data Sheet (SPA) STAT3 P-Y705 Kit / 62AT3PEG
62AT3PEG - Safety Data Sheet
Safety Data Sheet (ENG-GB) STAT3 P-Y705 Kit / 62AT3PEG
62AT3PEG - Safety Data Sheet
Safety Data Sheet (ENG-US) STAT3 P-Y705 Kit / 62AT3PEG
62AT3PEG - Safety Data Sheet
Safety Data Sheet STAT3 P-Y705 Kit / 62AT3PEH
62AT3PEH - Safety Data Sheet
Safety Data Sheet (DEU) STAT3 P-Y705 Kit / 62AT3PEH
62AT3PEH - Safety Data Sheet
Safety Data Sheet (ELL) STAT3 P-Y705 Kit / 62AT3PEH
62AT3PEH - Safety Data Sheet
Safety Data Sheet (FRA-FR) STAT3 P-Y705 Kit / 62AT3PEH
62AT3PEH - Safety Data Sheet
Safety Data Sheet (ITA) STAT3 P-Y705 Kit / 62AT3PEH
62AT3PEH - Safety Data Sheet
Safety Data Sheet (SPA) STAT3 P-Y705 Kit / 62AT3PEH
62AT3PEH - Safety Data Sheet
Safety Data Sheet (ENG-GB) STAT3 P-Y705 Kit / 62AT3PEH
62AT3PEH - Safety Data Sheet
Safety Data Sheet (ENG-US) STAT3 P-Y705 Kit / 62AT3PEH
62AT3PEH - Safety Data Sheet
Safety Data Sheet (DEU) STAT3 P-Y705 Kit / 62AT3PET
62AT3PET - Safety Data Sheet
Safety Data Sheet (ELL) STAT3 P-Y705 Kit / 62AT3PET
62AT3PET - Safety Data Sheet
Safety Data Sheet (FRA-FR) STAT3 P-Y705 Kit / 62AT3PET
62AT3PET - Safety Data Sheet
Safety Data Sheet (ITA) STAT3 P-Y705 Kit / 62AT3PET
62AT3PET - Safety Data Sheet
Safety Data Sheet (SPA) STAT3 P-Y705 Kit / 62AT3PET
62AT3PET - Safety Data Sheet
Safety Data Sheet (ENG-GB) STAT3 P-Y705 Kit / 62AT3PET
62AT3PET - Safety Data Sheet
Safety Data Sheet (ENG-US) STAT3 P-Y705 Kit / 62AT3PET
62AT3PET - Safety Data Sheet
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