Phospho-c-Jun (Ser63) cellular kit
Sensitive and robust assay to detect and study c-Jun
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This HTRF kit enables both SAPK/JNK and p38 MAPK pathway readout, measuring ATF2 modulation phosphorylated at Threonine-71.
ATF2 (Activating transcription factor 2) is a transcription factor activated by phosphorylation on threonin residues by JNK/SAPK. The phospho-ATF2 (Thr71) kit enables the quantitative detection of transcription factor ATF2 modulation phosphorylated at Threonine-71. ATF2 is also involved in PI3K/AKT, TNF, and estrogen signaling along with insulin secretion, thyroid hormone release, substance dependence, viral carcinogenesis and infectious diseases. The simple add-and-read protocol features no wash steps for a faster analysis in biological applications like oncology, diabetes or inflammation.
Mouse NIH3T3 cells were seeded at different cell densities in 96-well microplates, then stimulated with increasing concentrations of anisomycin for 30 minutes. Following the 2-plate assay protocol, 16 µL of lysate were transferred into a 384-well low volume white microplate before the addition of 4 µL of the HTRF phospho-ATF2 (Thr71) detection reagents. The HTRF signal was recorded after an overnight incubation.
Human HEK293A suspension cells were seeded at 100,000 cells/well in a 96 well half area plate, and incubated for 24h at 37°C, 5% CO2. Then cells were stimulated with different concentrations of anisomycin for 15, 30 & 60 minutes. After lysis, 16 µL of lysate were transferred into a 384-well low volume white microplate and 4 µL of the HTRF phospho-ATF2 (Thr71) detection reagents were added. The HTRF signal was recorded after an overnight incubation.
The human NIH3T3 cell line was seeded in a T175 flask and incubated a 37°C, 5% CO2 until confluency.
Serial dilutions of the cell lysate were performed in the supplemented lysis buffer, and 16 µL of each dilution were transferred into a low volume white microplate before the addition of 4 µL of HTRF phospho-ATF2 detection reagents. Equal amounts of lysates were used for a side by side comparison between HTRF and Western Blot.
Using the HTRF Phospho-ATF2 T71 assay, 3,000 cells/well were sufficient to detect a signal while 6,125 cells were needed using Western Blot with an ECL detection. These results demonstrate that the HTRF phospho-ATF2 assay is 2 times more sensitive than the Western Blot.
In response to cellular stress, such as exposure to genotoxic agents, cytokines, or UV irradiation, SAPK/JNK and p38 MAP kinases are activated and phosphorylate the transcription factor ATF-2 on residues Thr69 and Thr71. Once in the nucleus, phospho-ATF2 binds to cAMP response element (CRE) or to Activator Protein-1 (AP-1) response elements, and regulates the transcription of genes involved in cell growth, survival, and DNA damage response.
Simplified pathway dissection with HTRF phospho-assays and CyBi-felix liquid handling
Analyse of PI3K/AKT/mTor translational control pathway - Notes d'application
Cell-based kinase assays in HTS ? potential and limitations for primary and secondary screening
In collaboration with Bayer - Présentations scientifiaues
Cisbio lysis buffer compatibility
Cell Signaling: Biomarkers, Phospho- & total-protein Assays - Flyers
HTRF cellular phospho-protein assays
physiologically relevant results fo fast flowing research - 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
From 2D, 3D cell cultures to xenografts: A smart HTRF platform to maximize anticancer drug discovery
One technology across all samples - Notes d'application
HTRF phospho assays reveal subtle drug induced effects in tumor-xenografts
Tumor xenograft analysis: HTRF versus Western blot - Notes d'application
HTRF cell-based phospho-protein data normalization
Valuable guidelines for efficiently analyzing and interpreting results - Notes d'application
HTRF phospho-total lysis buffer: a universal alternative to RIPA lysis buffers
Increased flexibility of phospho-assays - Notes d'application
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 - Notes techniques
HTRF Alpha-tubulin Housekeeping kit
Properly interpret your compound effect - Notes d'application
Optimize your HTRF cell signaling assays on tissues
HTRF and WB compatible guidelines - Notes techniques
Key guidelines to successful cell signaling experiments
Mastering the art of cell signaling assays optimization - Guides
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 - Notes techniques
How to run a cell based phospho HTRF assay
What to expect at the bench - Vidéos
Unleash the potential of your phosphorylation research with HTRF
Unmatched ease of use, sensitivity and specificity assays - Vidéos
Product Insert ATF2 P-T71 Kit / 63ADK015PEG-63ADK015PEH
63ADK015PEG-63ADK015PEH - Protocoles
All your HTRF assays in one document! - Catalogue
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
HTRF® cell signaling platform combined with iCell® Hepatocytes
A solution for phospho-protein analysis in metabolic disorders - Posters
Unleash the potential of your phosphorylation research with HTRF
A fun video introducing you to phosphorylation assays with HTRF - Vidéos
How to run a cell based phospho HTRF assay
3' video to set up your Phospho assay - Vidéos
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. - Notes techniques
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 - Notes d'application
Methodological Aspects of Homogeneous Time-Resolved Fluorescence (HTRF)
Learn how to reduce time and sample consumption - Notes d'application
Safety Data Sheet ATF2 P-T71 Kit / 63ADK015PEG
63ADK015PEG - Fiches de sécurité
Safety Data Sheet (DEU) ATF2 P-T71 Kit / 63ADK015PEG
63ADK015PEG - Fiches de sécurité
Safety Data Sheet (ELL) ATF2 P-T71 Kit / 63ADK015PEG
63ADK015PEG - Fiches de sécurité
Safety Data Sheet (FRA-FR) ATF2 P-T71 Kit / 63ADK015PEG
63ADK015PEG - Fiches de sécurité
Safety Data Sheet (ITA) ATF2 P-T71 Kit / 63ADK015PEG
63ADK015PEG - Fiches de sécurité
Safety Data Sheet (SPA) ATF2 P-T71 Kit / 63ADK015PEG
63ADK015PEG - Fiches de sécurité
Safety Data Sheet (ENG-GB) ATF2 P-T71 Kit / 63ADK015PEG
63ADK015PEG - Fiches de sécurité
Safety Data Sheet (ENG-US) ATF2 P-T71 Kit / 63ADK015PEG
63ADK015PEG - Fiches de sécurité
Safety Data Sheet ATF2 P-T71 Kit / 63ADK015PEH
63ADK015PEH - Fiches de sécurité
Safety Data Sheet (DEU) ATF2 P-T71 Kit / 63ADK015PEH
63ADK015PEH - Fiches de sécurité
Safety Data Sheet (ELL) ATF2 P-T71 Kit / 63ADK015PEH
63ADK015PEH - Fiches de sécurité
Safety Data Sheet (FRA-FR) ATF2 P-T71 Kit / 63ADK015PEH
63ADK015PEH - Fiches de sécurité
Safety Data Sheet (ITA) ATF2 P-T71 Kit / 63ADK015PEH
63ADK015PEH - Fiches de sécurité
Safety Data Sheet (SPA) ATF2 P-T71 Kit / 63ADK015PEH
63ADK015PEH - Fiches de sécurité
Safety Data Sheet (ENG-GB) ATF2 P-T71 Kit / 63ADK015PEH
63ADK015PEH - Fiches de sécurité
Safety Data Sheet (ENG-US) ATF2 P-T71 Kit / 63ADK015PEH
63ADK015PEH - Fiches de sécurité
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