Phospho-BAD (Ser112) cellular kit
Highly adaptable assay to quantify phospho-BAD (Ser112)
Phospho-mTOR (Ser2448) cellular kit HTRF®
The phospho-mTOR (Ser2448) kit enables the cell-based quantitative detection of phosphorylated mTOR.
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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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No-wash
Overview
The Phospho-mTOR (Ser2448) cellular assay kit is ideal for measuring phosphorylated mTOR directly in cells. Using a streamlined protocol, amenable to low-volume formats, this kit can be used from basic research to High Throughput drug screening.
Benefits
- COMPATIBLE WITH MANY CELL TYPES
- PRECISION
Phospho-mTor (Ser2448) assay principle
The Phospho-mTor (Ser2448) assay measures mTor when phosphorylated at Ser2448. Contrary to Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis or transfer. The Phospho-mTor (Ser2448) 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-mTor (Ser2448) 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-mTor (Ser2448) HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.
Phospho-mTor (Ser2448) 1-plate assay protocol
Detection of Phosphorylated mTor (Ser2448) 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 assay compared to WB using phospho-mTOR cellular assay
Hek293 cells were grown in a T175 flask 37°C, 5% Co2, 2 days.After elimination of cell culture medium, 3 mL of supplemented lysis buffer were added and incubated for 30 min.Soluble supernatants were collected after 10 min centrifuging. Equal amounts of lysates were used for a side by side comparison of WB and HTRF.
HTRF assay shows the same level of sensitivity as Western Blot: 1560 cells for the two technologies.
Insulin dose-response on NIH 3T3 cells
Murine NIH 3T3 cells (100,000 cells/well) were stimulated for 30 minutess at 37°C with various concentrations of forskolin. After a 30 minutes lysis incubation time, phosphorylated mTOR was measured using the two-plate assay protocol.
Inhibition effect of Rapamycin on basal level of Jurkat cells
Jurkat cells (100,000 cells/well) were incubated for 30 min at 37°C with various concentrations of inhibitor. After a 30 minutes lysis incubation time, inhibition of the basal level of phosphorylated mTOR was measured using the one-plate assay protocol.
Simplified Pathway for Phospho-mTor (Ser2448) assay kit
The mammalian target of rapamycin (mTOR) is a Ser/Thr protein kinase that functions as an ATP and amino acid sensor and plays a key role in cell growth and homeostasis. As mTOR may be abnormally regulated in tumors, it is currrently under investigation as a potential target for anti-cancer therapy. mTOR is itself phosphorylated at Ser2448 via the PI3 kinase/Akt signaling pathway, and autophosphorylated at Ser2481.
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
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 and Alpha 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
Cell-based kinase assays in HTS ? potential and limitations for primary and secondary screening
In collaboration with Bayer - Scientific Presentations
Manual mTOR P-S2448 Kit / 64TORPEG-64TORPEH
64TORPEG-64TORPEH - Product Insert
All your HTRF assays in one document! - Catalog
Immuno-Oncology Drug Discovery
Explore our solutions for Immuno-oncology research - Flyers
An innate and adaptive immunity recap
Insight into the diversity of cells & signaling pathways - Guides
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
How HTRF compares to Western Blot and ELISA
Get the brochure about technology comparison. - Brochures
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 (DEU) mTOR P-S2448 Kit / 64TORPEG
64TORPEG - Safety Data Sheet
Safety Data Sheet (ELL) mTOR P-S2448 Kit / 64TORPEG
64TORPEG - Safety Data Sheet
Safety Data Sheet (FRA-FR) mTOR P-S2448 Kit / 64TORPEG
64TORPEG - Safety Data Sheet
Safety Data Sheet (ITA) mTOR P-S2448 Kit / 64TORPEG
64TORPEG - Safety Data Sheet
Safety Data Sheet (SPA) mTOR P-S2448 Kit / 64TORPEG
64TORPEG - Safety Data Sheet
Safety Data Sheet (ENG-GB) mTOR P-S2448 Kit / 64TORPEG
64TORPEG - Safety Data Sheet
Safety Data Sheet (ENG-US) mTOR P-S2448 Kit / 64TORPEG
64TORPEG - Safety Data Sheet
Certificate of Analysis mTOR P-S2448 Kit / 64TORPEG
64TORPEG Batch 04D - Quality Control Report
Safety Data Sheet (DEU) mTOR P-S2448 Kit / 64TORPEH
64TORPEH - Safety Data Sheet
Safety Data Sheet (ELL) mTOR P-S2448 Kit / 64TORPEH
64TORPEH - Safety Data Sheet
Safety Data Sheet (FRA-FR) mTOR P-S2448 Kit / 64TORPEH
64TORPEH - Safety Data Sheet
Safety Data Sheet (ITA) mTOR P-S2448 Kit / 64TORPEH
64TORPEH - Safety Data Sheet
Safety Data Sheet (SPA) mTOR P-S2448 Kit / 64TORPEH
64TORPEH - Safety Data Sheet
Safety Data Sheet (ENG-GB) mTOR P-S2448 Kit / 64TORPEH
64TORPEH - Safety Data Sheet
Safety Data Sheet (ENG-US) mTOR P-S2448 Kit / 64TORPEH
64TORPEH - Safety Data Sheet
Safety Data Sheet (DEU) mTOR P-S2448 Kit / 64TORPEY
64TORPEY - Safety Data Sheet
Safety Data Sheet (ELL) mTOR P-S2448 Kit / 64TORPEY
64TORPEY - Safety Data Sheet
Safety Data Sheet (FRA-FR) mTOR P-S2448 Kit / 64TORPEY
64TORPEY - Safety Data Sheet
Safety Data Sheet (ITA) mTOR P-S2448 Kit / 64TORPEY
64TORPEY - Safety Data Sheet
Safety Data Sheet (SPA) mTOR P-S2448 Kit / 64TORPEY
64TORPEY - Safety Data Sheet
Safety Data Sheet (ENG-GB) mTOR P-S2448 Kit / 64TORPEY
64TORPEY - Safety Data Sheet
Safety Data Sheet (ENG-US) mTOR P-S2448 Kit / 64TORPEY
64TORPEY - Safety Data Sheet
Plate Reader Requirement
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