Powerful and most sensitive HTS plate reader
The PHERAstar® FSX is our latest, most sensitive multi-mode reader for high-throughput screening, combining this sensitivity with fast read times. Thanks to the unique Optic Module system, Simultaneous Dual Emission, UV/vis spectrometer, and dedicated AlphaScreen® and TRF lasers, the PHERAstar FSX will perform your assay with ease and speed, even in 3456-well microplates.
No longer worry about installing filters and dichroics: the PHERAstar FSX comes with easy-to-handle, assay-optimized Optic Modules which contain all application-specific filters, mirrors, dichroics and/or polarizers, and are automatically recognized by the plate reader. In addition, the PHERAstar FSX is equipped with four matched and assay-optimized photomultiplier tubes (PMTs) which are automatically selected for the relevant detection mode. You can also measure assays with two emission wavelengths at the same time thanks to the Simultaneous Dual Emission.
For HTS automation purposes, the PHERAstar FSX offers improved robotic integration capabilities and multi-user control.
- The most sensitive reader in fluorescence intensity and polarization
- The only plate reader with 5 assay-dedicated detectors
- Simultaneous Dual Emission, including for Alpha Technology
- 9 decades luminescence dynamic range
- Dedicated AlphaScreen®/AlphaLISA®/AlphaPlex™ laser
- New generation TRF-laser
- Top and bottom focal height adjustment
- High-precision injectors with simultaneous reagent injection and detection
- High-speed absorbance measurements
PHERAstar measures AlphaScreen assay to develop selective inhibitors for the human YEATS domainsThomas Christott , Carmen Coxon , James Bennett , Charline Giroud , Octovia Monteiro , Oleg Fedorov , , Structural Genomics Consortium, University of Oxford, UK, 04/2018
YEATS domains are epigenetic regulators and are recognised as readers of histone post-translational modifications (HPTM) alongside bromodomains, PHD fingers, and others. YEATS domains bind to lysine when the ε-carbon is acetylated or crotonylated. The YEATS-containing ENL links histone acetylation to active transcription and is a major driver of several types of acute leukaemia. Hence, ENL is a rational drug target to attenuate aberrant cell growth and malignancy.
An AlphaScreen assay that reports on the interaction of modified histone 3 with the YEATS domains of different proteins such as ENL was developed. The inhibitor screening was performed on a PHERAstar® microplate reader and uncovered a potent small molecule inhibitor interfering with the YEATS domains of ENL and AF9.
The PHERAstar FSX provided a powerful and versatile platform for the drug discovery campaign. Here, the inhibitor screening of tens of thousands of compounds for inhibiting the YEATS domain with an AlphaScreen® approach resulted in the identification of a potent small molecule inhibitor.
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.