Phospho-ATF2 (Thr71) cellular kit
Phospho-c-Jun (Ser63) cellular kit HTRF®
The Phospho-c-Jun (Ser63) kit enables the cell-based quantitative detection of cJun protein phosphorylated at Ser63.
-
Ease-of-use
-
High sensitivity
Overview
The Phospho-c-Jun (Ser63) Cellular Assay kit is designed for the robust quantification of phoshorylated c-Jun at Ser-36 directly in cell lysates. Using a streamlined mix-and-read no-wash protocol, this kit can be used from basic research to High Throughput drug screening.
Benefits
- COMPATIBLE WITH MANY CELL TYPES
- SPECIFICITY
Phospho-c-Jun (Ser63) assay principle
The Phospho-c-Jun (Ser63) assay measures c-Jun when phosphorylated at Ser63. Contrary to Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis or transfer. The Phospho-c-Jun (Ser63) 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-c-Jun (Ser63) 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-c-Jun (Ser63) HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.
Phospho-c-Jun (Ser63) 1-plate assay protocol
Detection of Phosphorylated c-Jun (Ser63) 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.
WB versus HTRF assay for Phoshpo-c-Jun (Ser63) kit
HEK293 cells were cultured for two days and stimulated with 500nM anisomycin for 30 min. After removal of cell culture medium, lysis buffer was added and the cells were incubated for 30 min. Supernatants were collected following centrifugation and equal amounts of lysates were used for a side by side comparison of WB and HTRF. The HTRF assay proved to be 8-fold more sensitive than the western-blot assay.
Anysomycin dose-response on NIH-3T3 cells for Phospho-c-Jun kit
Murine NIH-3T3 cells (50,000 cells/well) were stimulated for 30 minutess at 37°C with various concentrations of anisomycin. After a 30 minutes lysis incubation time, phosphorylated c-Jun was measured using the two-plate assay protocol of the Phospho-c-Jun assay.
Inhibition effect of c-Jun on HEK293 stimulated with anisomycin
HEK293 cells (50,000 cells/well) were incubated for 1h at 37°C with various concentrations of cell-permeable c-Jun peptide. 50nM anisomycin (agonist) were then added and incubated for 30 minutess. After a 30 minutes lysis incubation time, inhibition of c-Jun phosphorylation was measured using the Phospho-c-Jun assay. Assay was done using the two-plate assay protocol.
Simplified pathway of c-Jun protein family
c-Jun belongs to the mammalian Jun protein family which are transcriptoin factors that have to dimerize to bind to their target DNA sites. c-Jun is a major component of the Activator Protein 1 (AP-1) complex. Phosphorylation of c-Jun at Ser63 and Ser73 through JNK stimulates AP-1. Once phosphorylated, c-Jun can dimerize and bind to DNA sequences and affect transription. c-Jun regulates numerous cellular responses including proliferation, malignant transformation, differentiation and apoptosis.
Simplified pathway dissection with HTRF phospho-assays and CyBi-felix liquid handling
Analyse of PI3K/AKT/mTor translational control pathway - Application Notes
Cell-based kinase assays in HTS ? potential and limitations for primary and secondary screening
In collaboration with Bayer - 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
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 - 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
Product Insert cJun P-S63 Kit / 64JUNPEG-64JUNPEH
64JUNPEG-64JUNPEH - 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
An innate and adaptive immunity recap
Insight into the diversity of cells & signaling pathways - Guides
How to run a cell based phospho HTRF assay
3' video to set up your Phospho assay - Videos
Unleash the potential of your phosphorylation research with HTRF
A fun video introducing you to phosphorylation assays with HTRF - Videos
Advance your research on Fibrosis
Kits and reagents for Fibrosis 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 (DEU) cJun P-S63 Kit / 64JUNPEG
64JUNPEG - Safety Data Sheet
Safety Data Sheet (ELL) cJun P-S63 Kit / 64JUNPEG
64JUNPEG - Safety Data Sheet
Safety Data Sheet (FRA-FR) cJun P-S63 Kit / 64JUNPEG
64JUNPEG - Safety Data Sheet
Safety Data Sheet (ITA) cJun P-S63 Kit / 64JUNPEG
64JUNPEG - Safety Data Sheet
Safety Data Sheet (SPA) cJun P-S63 Kit / 64JUNPEG
64JUNPEG - Safety Data Sheet
Safety Data Sheet (ENG-GB) cJun P-S63 Kit / 64JUNPEG
64JUNPEG - Safety Data Sheet
Safety Data Sheet (ENG-US) cJun P-S63 Kit / 64JUNPEG
64JUNPEG - Safety Data Sheet
Safety Data Sheet (DEU) cJun P-S63 Kit / 64JUNPEH
64JUNPEH - Safety Data Sheet
Safety Data Sheet (ELL) cJun P-S63 Kit / 64JUNPEH
64JUNPEH - Safety Data Sheet
Safety Data Sheet (FRA-FR) cJun P-S63 Kit / 64JUNPEH
64JUNPEH - Safety Data Sheet
Safety Data Sheet (ITA) cJun P-S63 Kit / 64JUNPEH
64JUNPEH - Safety Data Sheet
Safety Data Sheet (SPA) cJun P-S63 Kit / 64JUNPEH
64JUNPEH - Safety Data Sheet
Safety Data Sheet (ENG-GB) cJun P-S63 Kit / 64JUNPEH
64JUNPEH - Safety Data Sheet
Safety Data Sheet (ENG-US) cJun P-S63 Kit / 64JUNPEH
64JUNPEH - Safety Data Sheet
Plate Reader Requirement
Choosing the right microplate reader ensures you’ll get an optimal readout. Discover our high performance reader, or verify if your lab equipment is going to be compatible with this detection technology.
Let's find your reader
You might be interested in