Lector de placas HTS potente y sensible
El PHERAstar FSX es el lector más novedoso y rápido para High-Throughput Screening, combina su altísima sensibilidad con rapidísimos tiempos de lectura. Gracias a sus exclusivo sistema de Módulo Óptico, Emisión Simultanea Dual, Espectrómetro UV/Vis, y láseres dedicados para Alphascreen y TRF, cualquiera que sea su aplicación, el PHERAstar FSX le realizará fácil y rápidamente, icluso en placas de 3456 pocillos.
No se preocupe por que filtro o dicróico esté instalado en su PHERAstar FSX. Los Módulos Ópticos son muy fáciles de manejar y contienen los filtros, espejos, dicróicos y/o polarizadores para cada aplicación y son reconocidos automáticamente por el lector. Además el PHERAstar FSX está equipado con 4 tubos fotomultiplicadores (PMTs) que son seleccionados automáticamente para cada modo de detección, así como para Emisión Simultanea Dual.
Para la automatización para HTS el PHERAstar FSX ofrece capacidades de integración robótica mejoradas y control multi-usuari.
- El lector más sensible en Intensidad de Fluorescencia y Fluorescencia Polarizada
- El único lector de placas con 5 detectores dedicados al ensayo
- Emisión dual simultánea, incluida la tecnología Alpha
- Rango dinámico de 9 décadas en luminiscencia
- Laser dedicado para AlphaScreen®/AlphaLISA®/Emisión Simultanea Dual Alpha PlexTM
- Laser de TRF de última generación
- Lectura superior e inferior con ajuste automático de altura focal
- inyectores de alta precisión permitiendo inyección y lectura simultanea
- Mediciones de absorbancia de alta velocidad
Cell-based assay detects residual β-blocker substances in effluent of municipal wastewater treatment plantsK. Bernhard , C. Stahl , R. Martens , M. Frey, SIZ Zellkulturtechnik, c/o Hochschule Mannheim, Mannheim, Germany, 03/2018
Residues of human pharmaceuticals, such as β-blockers are increasingly found in effluent wastewater of treatment plants (WWTP) and represent a potential environmental threat. Beta-blockers antagonize β-adrenergic receptors and thereby control hypertension and cardiac arrhythmias. The amino acid sequence of the target, the β1-adrenergic receptor, is evolutionarily highly conserved among vertebrates. Thus, organisms may physiologically respond to β-blockers. For environmental risk assessment one has to know the extent to which aquatic organisms are exposed to β-blockers and metabolites with the same mode of action (MOA).
The mode of action-based β-blocker assay in living cells measures total β-blocker activities in complex mixtures such as WWTP effluents as equivalents of the lead substance metoprolol (MetEQ). The assay is suitable as a standard method in large-scale monitoring of WWTP effluents and the aquatic environment they are discharged into. The CLARIOstar® microplate reader provides highly stable and reliable results as well as the required robustness for continuous use.
FRET sensor assesses ATP levels in living plants experiencing low oxygen conditionsStephan Wagner1,2 , Philippe Fuchs1,2 , Thomas Nietzel1,2 , Marlene Elsässer1,2 , Markus Schwarzländer1,2, 1 Institute of Plant Biology and Biotechnology, University of Münster, Germany , 2 Institute of Crop Science and Resource Conservation (INRES), University of Bonn, Germany, 02/2018
To survive as sessile organisms, plants need to constantly adapt their metabolism to their environment. Flooding of plants is widespread and has severe metabolic consequences as it limits the cellular supply with oxygen to drive respiration, and thus production of ATP. Submergence acclimation typically involves drastic alterations in metabolism to circumvent anoxia and to maintain primary metabolism and ATP supply. We have recently established the use of a genetically encoded protein sensor (ATeam1.03-nD/nA) to assess MgATP dynamics in the model plant Arabidopsis thaliana.
This FRET-system read on a CLARIOstar® allows for reliable analyses of changes in MgATP in vivo and in real-time and provides a new handle to investigate the energetic consequences of low oxygen for the plant cell. The microplate reader’s orbital averaging function acquires the signal in multiple points of the well to obtain a meaningful result of the non-homogenously distributed Arabidopsis seedlings.
NanoBRET™ assay quantitatively evaluates VEGF binding to the VEGFR2 in real-time in living cellsKilpatrick LE1 , Friedman-Ohana R2 , Alcobia DC1 , Riching K2 , Peach CJ1 , Wheal AJ1 , Briddon SJ1 , Robers MB2 , Zimmerman K2 , Machleidt T2, , Wood KV2 , Woolard J1 , Hill SJ1, 1Division of Physiology, Pharmacology and Neuroscience, University of Nottingham, UK , 2Promega Corporation, Madison, WI, USA, 01/2018
Receptor tyrosine kinases (RTK) are transmembrane receptors that translate stimulation by growth factors into regulation of cell growth, proliferation, cell death and differentiation. Consequently, aberrant RTK-signaling is implicated in cancer, making it a popular anti-cancer drug target. Development of pharmaceutical RTK-inhibitors requires assays that characterize receptor-ligand interaction and that identify inhibitors thereof.
A novel method labeled the vascular endothelial growth factor (VEGF) at a single site with the fluorophore TMR. The labeled ligand was used in combination with HEK293 cells expressing the RTK VEGF-receptor 2 (VEGFR2) fused to the NanoLuc® luciferase. Upon interaction of ligand and receptor, bioluminescence resonance energy transfer (BRET) between luciferase and fluorophore takes place.
The signal of luciferase and fluorophore were acquired simultaneously with the PHERAstar microplate reader. This way, not only the pKi of a VEGF-inhibitor was determined, but also the time-course of binding and inhibition was monitored.
CRISPR/Cas9 genome-edited cells express nanoBRET-donor that monitors protein interaction and traffickingCarl White1,2 , Ethan See1,2 , Kevin Pfleger1,2,3, 1Molecular Endocrinology and Pharmacology, Harry Perkins Institute of Medical Research, Australia , 2Centre for Medical Research, The University of Western Australia, Australia , 3Dimerix Limited, Nedlands, Australia, 01/2018
GPCRs are important drug targets requiring receptor-protein interaction and trafficking studies to reveal how they function. Bioluminescence resonance energy transfer (BRET) is a versatile tool to study such interactions and trafficking. Hitherto, it is limited by the ectopic expression of labelled interaction partners. CRISPR/Cas9 genome editing overcomes the limitation by enabling endogenous expression of luciferase-labelled proteins.
CRISPR/Cas9-edited cells endogenously expressing a CXCR4/NanoLuciferase fusion protein were used in conjunction with β-Arrestin/Venus to monitor receptor activation. Employing two fluorophores fused to a membrane and endosome standing CXCR4-interacting protein, respectively, allowed for monitoring of receptor trafficking.
The novel CRISPR/Cas9 technique successfully fused the Nluc BRET donor to endogenously-expressed CXCR4. The resulting protein levels were sufficient to monitor receptor interactions as well as internalization. The internalization assay depends on two acceptor fluorophores whose selective detection was rendered possible by the CLARIOstar’s monochromator.
Assay development for essential enzyme activity in the tegument of live SchistosomesMadhu Sundaraneedi1 , Luke Becker2 , Giovanni Abbenante3 , Alex Loukas2 , Grant Collins1 , Mark Pearson2, 1School of Physical, Environmental and Mathematical Sciences, University of New South Wales, Australia , 2Australian Institute for Tropical Health and Medicine, James Cook University, Australia , 3BMG LABTECH Australia, 12/2017
Schistosomiasis is a parasitic disease that affects over 200 million people in tropical, developing nations, causing severe morbidity and over 300,000 deaths annually. Schistosomiasis is treated with a single drug and no vaccine is available.
We selected three Schistosoma surface-associated enzymes that are indispensable to parasitic survival: alkaline phosphatase; phosphodiesterase SmNPP-5 and an acetylcholinesterase. The activity of these molecules on the surface of live and intact larval and adult Schistosoma can be assayed in real-time of cultured parasites, providing a tool to assess the efficacy of drugs or vaccines targeting these enzymes. The colorimetric assay was read on a FLUOstar® Omega microplate reader.
The versatile instrument supports development of assays and drugs. Apart from its absorbance spectrometer, the FLUOstar Omega is equipped with fluorescence and luminescence detection capabilities allowing fast and reliable endpoint and kinetic measurements in all detection modes.