Advanced phospho-ERK (Thr202/Tyr204) cellular kit
Simple, all-in-one kit for robust detection of Phospho-ERK.
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The phospho-HER3 (Tyr1289) kit enables the cell-based detection of human epidermoid growth factor 3 (HR3, Erbb3) signaling activity when phosphorylated on Tyr1289
The phospho-HER3 (Tyr1289) assay is designed for the robust quantification of HER3 (Erbb3) modulation when phosphorylated on Tyr1289, an indicator of many types of cancer pathologies. Overexpression of HER3, or human epidermoid receptor 3/ ErbB3, is considered as an indicator for a poor prognosis in human breast cancer. Heterodimerization of HER3 with HER2 is associated with an increase in the aggressiveness of a breast cancer. HER3 is also been found to be involved in prostate, gastric, colon, or other carcinomas. This makes HER3 a key target for anti-cancer therapies.
Breast cancer cells were grown in a T175 flask at 37 °C, 5% CO2 for 48 h. Cells were then stimulated with 100 nM HRG Beta1 for 10 min. After medium removal, the cells were lysed with 3 mL of supplemented lysis buffer for 30 min at room temperature. Soluble fractions were collected after a 10 min centrifugation. Serial dilutions of the cell lysate were performed in the supplemented lysis buffer and 16 µL of each dilution were dispensed and analyzed side-by-side by Western Blot and by HTRF. Using HTRF phospho-HER3 (Tyr1289) only 5,000 cells are sufficient for minimal signal detection, while 80,000 cells are needed for a Western Blot signal. The HTRF phospho-HER3 assay is at least 16-fold more sensitive than the Western Blot.
100,000 human recombinant HER2/HER3, MCF-7, SK-BR-3 and breast cancer cells were plated in 96 well plates and incubated for 24h at 37 °C-5% Co2. After incubation with increasing concentrations of HRG Beta3 (10min), medium was removed and cells were lysed with 50µl of Lysis buffer for 30min at RT under gentle shaking. 16 µL of lysate were transferred into a 384-well sv white microplate and 4 µL of the HTRF phospho-HER3 detection reagents were added. HTRF signal was recorded after an overnight incubation.
Human breast carcinoma and SKBR3 cells were pre-treated for 30 min with a dose-response of Lapatinib, a small tyrosine kinase inhibitor, then stimulated for 10 min with 100 nM HRG Beta1 for 10 min.
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
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
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
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
A guide to Homogeneous Time Resolved Fluorescence
General principles of HTRF - Guides
How to run a cell based phospho HTRF assay
3' video to set up your Phospho assay - Videos
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
Manual HER3 P-Y1289 Kit / 64HR3PEG-64HR3PEH
64HR3PEG-64HR3PEH - Product Insert
Certificate of Analysis HER3 P-Y1289 Kit / 64HR3PEG
64HR3PEG Batch 03A - Quality Control Report
Safety Data Sheet (DEU) HER3 P-Y1289 Kit / 64HR3PEG
64HR3PEG - Safety Data Sheet
Safety Data Sheet (ELL) HER3 P-Y1289 Kit / 64HR3PEG
64HR3PEG - Safety Data Sheet
Safety Data Sheet (FRA-FR) HER3 P-Y1289 Kit / 64HR3PEG
64HR3PEG - Safety Data Sheet
Safety Data Sheet (ITA) HER3 P-Y1289 Kit / 64HR3PEG
64HR3PEG - Safety Data Sheet
Safety Data Sheet (SPA) HER3 P-Y1289 Kit / 64HR3PEG
64HR3PEG - Safety Data Sheet
Safety Data Sheet (ENG-GB) HER3 P-Y1289 Kit / 64HR3PEG
64HR3PEG - Safety Data Sheet
Safety Data Sheet (ENG-US) HER3 P-Y1289 Kit / 64HR3PEG
64HR3PEG - Safety Data Sheet
Safety Data Sheet (DEU) HER3 P-Y1289 Kit / 64HR3PEH
64HR3PEH - Safety Data Sheet
Safety Data Sheet (ELL) HER3 P-Y1289 Kit / 64HR3PEH
64HR3PEH - Safety Data Sheet
Safety Data Sheet (FRA-FR) HER3 P-Y1289 Kit / 64HR3PEH
64HR3PEH - Safety Data Sheet
Safety Data Sheet (ITA) HER3 P-Y1289 Kit / 64HR3PEH
64HR3PEH - Safety Data Sheet
Safety Data Sheet (SPA) HER3 P-Y1289 Kit / 64HR3PEH
64HR3PEH - Safety Data Sheet
Safety Data Sheet (ENG-GB) HER3 P-Y1289 Kit / 64HR3PEH
64HR3PEH - Safety Data Sheet
Safety Data Sheet (ENG-US) HER3 P-Y1289 Kit / 64HR3PEH
64HR3PEH - Safety Data Sheet
Safety Data Sheet (DEU) HER3 P-Y1289 Kit / 64HR3PEY
64HR3PEY - Safety Data Sheet
Safety Data Sheet (ELL) HER3 P-Y1289 Kit / 64HR3PEY
64HR3PEY - Safety Data Sheet
Safety Data Sheet (FRA-FR) HER3 P-Y1289 Kit / 64HR3PEY
64HR3PEY - Safety Data Sheet
Safety Data Sheet (ITA) HER3 P-Y1289 Kit / 64HR3PEY
64HR3PEY - Safety Data Sheet
Safety Data Sheet (SPA) HER3 P-Y1289 Kit / 64HR3PEY
64HR3PEY - Safety Data Sheet
Safety Data Sheet (ENG-GB) HER3 P-Y1289 Kit / 64HR3PEY
64HR3PEY - Safety Data Sheet
Safety Data Sheet (ENG-US) HER3 P-Y1289 Kit / 64HR3PEY
64HR3PEY - Safety Data Sheet
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