cAMP Gs dynamic kit
Optimal performance when applied to Gs activation or inhibition
Phospho-Phospholamban (Thr17) cellular kit
The phospho-Phospholamban (Thr17) kit enables the cell-based quantitative detection of phosphorylated Phospholamban at Thr17 (CaMKII activation).
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Ready-to-use
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High sensitivity
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Low sample consumption
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No-wash
Overview
This cell-based assay enables the detection of Phosphorylated Phospholamban (PLN) at Threonine 17. The phospho assay along with the total PLN assay are ideal for monitoring the activation of CaMKII. The increase in intracellular Ca2+ causes the activation of CaMKII, which in turn phosphorylates PLN at the Thr17 residue. Dephosphorylated PLN inhibits SERCA2a, whereas phosphorylation of PLN at either the Ser16 site by PKA or the Thr17 site by CaMKII reverses this SERCA2a inhibition. The PLN phosphorylation state regulates the activity of this Ca2+ pump.
Benefits
- SPECIFICITY
- PRECISION
- DATA NORMALIZATION
Phospho-Phospholamban (Thr17) assay principle
The Phospho-Phospholamban (Thr17) assay measures Phospholamban when phosphorylated at Thr17. Contrary to Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis or transfer. The Phospho-Phospholamban (Thr17) 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-Phospholamban (Thr17) 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-Phospholamban (Thr17) HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.
Phospho-Phospholamban (Thr17)1-plate assay protocol
Detection of Phosphorylated Phospholamban (Thr17)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.
Phospho-phospholamban assay on heart homogenate
Spontaneously hypertensive (SHR) heart and normal heart samples were homogenized using Cisbio lysis buffer and supernatant was collected after centrifugation. 16 µL of lysate were transferred into a 384-well sv white microplate and 4 µL of the HTRF phospho-Phospholamban (Thr17) detection reagents were added.
The HTRF signal was recorded after an overnight incubation at room temperature.
Simplified pathway dissection with HTRF phospho-assays and CyBi-felix liquid handling
Analyse of PI3K/AKT/mTor translational control pathway - Application Notes
Open R&D: Sanofi Access Platform
In collaboration with Sanofi - Scientific Presentations
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
Save time and money
Switch to HTRF assays - Flyers
Species compatibility
Cell Signaling: Biomarkers, Phospho- & total-protein assays - Flyers
Side-by-side comparison of HTRF, Western Blot, ELISA and AlphaScreen® SureFire®
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
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
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
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 phospho-assays reveal subtle drug-induced effects
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
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 PLamban P-T17 Kit / 63ADK074PEG
63ADK074PEG - Product Insert
Product Insert PLamban P-T17 Kit / 63ADK074PEH
63ADK074PEH - Product Insert
Cisbio Product Catalog 2019
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
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 - Videos
How to run a cell based phospho HTRF assay
3' video to set up your Phospho assay - Videos
Safety Data Sheet (DEU-) PLamban P-T17 Kit / 63ADK074PEG
63ADK074PEG - Safety Data Sheet
Safety Data Sheet (ELL-) PLamban P-T17 Kit / 63ADK074PEG
63ADK074PEG - Safety Data Sheet
Safety Data Sheet (FRA-FR) PLamban P-T17 Kit / 63ADK074PEG
63ADK074PEG - Safety Data Sheet
Safety Data Sheet (ITA-) PLamban P-T17 Kit / 63ADK074PEG
63ADK074PEG - Safety Data Sheet
Safety Data Sheet (SPA-) PLamban P-T17 Kit / 63ADK074PEG
63ADK074PEG - Safety Data Sheet
Safety Data Sheet (ENG-GB) PLamban P-T17 Kit / 63ADK074PEG
63ADK074PEG - Safety Data Sheet
Safety Data Sheet (ENG-US) PLamban P-T17 Kit / 63ADK074PEG
63ADK074PEG - Safety Data Sheet
Safety Data Sheet (DEU-) PLamban P-T17 Kit / 63ADK074PEH
63ADK074PEH - Safety Data Sheet
Safety Data Sheet (ELL-) PLamban P-T17 Kit / 63ADK074PEH
63ADK074PEH - Safety Data Sheet
Safety Data Sheet (FRA-FR) PLamban P-T17 Kit / 63ADK074PEH
63ADK074PEH - Safety Data Sheet
Safety Data Sheet (ITA-) PLamban P-T17 Kit / 63ADK074PEH
63ADK074PEH - Safety Data Sheet
Safety Data Sheet (SPA-) PLamban P-T17 Kit / 63ADK074PEH
63ADK074PEH - Safety Data Sheet
Safety Data Sheet (ENG-GB) PLamban P-T17 Kit / 63ADK074PEH
63ADK074PEH - Safety Data Sheet
Safety Data Sheet (ENG-US) PLamban P-T17 Kit / 63ADK074PEH
63ADK074PEH - Safety Data Sheet