Most flexible plate reader with LVF monochromators
The CLARIOstar® is BMG LABTECH’s most flexible microplate reader, equipped with our revolutionary LVF monochromator™ technology. A combination of monochromators, filters, and spectrometer means that it does not compromise on sensitivity or flexibility. As well as its high performance in all detection modes, the CLARIOstar’s versatility also makes it ideal for assay development.
The CLARIOstar is a multi-mode microplate reader with advanced LVF Monochromators™, highly sensitive filters, and an ultra-fast UV/vis spectrometer. It is a modular microplate reader with up to seven different detection modes: fluorescence intensity (incl. FRET), fluorescence polarization, luminescence (including BRET), UV/vis absorbance, time-resolved fluorescence, TR-FRET, and AlphaScreen®/AlphaLISA®/AlphaPlex™.
- Innovative monochromators for increased sensitivity
- Fully-flexible wavelengths and bandwidths
- Spectral scanning in AS, FI and LUM
- Higher light transmission
- High-speed absorbance measurements
- Dedicated laser for Alpha Technology
- Live cell-based assays
- Low volume measurements
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.
Fluorescence Polarization based assay for rapid, precise, high-throughput measurement of IgG & Fc containing derivativesDr. Carolanne Doherty, Valitacell, NIBRT, Fosters Avenue, Blackrock, Dublin, Ireland, 11/2017
The accurate, rapid and high-throughput measurement of IgG is essential in the development and manufacture of most therapeutic antibodies. Monoclonal antibodies are becoming increasingly dominant in biopharmaceuticals, where a vast number of samples must be screened for the development of each potential therapeutic.
Here we report the development of a novel, rapid, and simple fluorescence polarization based assay for high-throughput titer measurement of IgG and Fc-containing derivatives. It uses fluorescently labeled protein G that binds IgG. Upon binding, the depolarization of emitted light decreases, this can be detected to report on the presence of IgG. The CLARIOstar®, PHERAstar® and POLARstar® exhibit excellent assay quality when used with the ValitaTITER assay, which enables a high-throughput, simple, precise method for quantification of IgG. Moreover, the assay can be performed using sample straight cell culture supernatant, which means there are no complex sample preparation steps.
Calcium retention capacity assay evaluates inhibition of mitochondrial permeability transition poreM. Awais , D. Latawiec , R. Sutton, Liverpool Pancreatitis Research Group, Institute of Translational Medicine, University of Liverpool, UK, 11/2017
Mitochondrial dysfunction is central to the pathogenesis of acute pancreatitis, ischemia-reperfusion injury of the heart, brain and kidney, muscular dystrophies and neurodegeneration. Mitochondrial dysfunction is the result of a sudden increase in permeability of the inner mitochondrial membrane (IMM), via persistent opening of a multi-protein channel known as the mitochondrial permeability transition pore (MPTP). This is followed by uncontrolled proton flow across the IMM and unregulated flux of water, ions and small solutes into and out of the mitochondrial matrix. This results in rupture of the outer mitochondrial membrane (OMM) and eventually cell death by necrosis. Therefore, MPTP is an attractive target for cell death prevention in a host of disease states.
The calcium retention capacity assay challenges isolated mitochondria with spikes of calcium ions. Upon opening of the MPTP, Ca2+ leaks into the assay buffer and increases fluorescence of the membrane-impermeable CalciumGreen™ dye. The Omega multi-mode plate reader has proven excellent robustness for performing the multiple injections as well as reliable fluorescent detection of the assay.
Protein-induced fluorescence enhancement detects protein-nucleic acid interactions in microplatesFulneček J. , Valuchová S. , Petrov A.P. , Tripsianes K. , Říha K., Research group Molecular Biology of Plants, Central European Institute of Technology, Masaryk University, Brno, Czech Republic, 10/2017
Various biological processes depend on the interaction of proteins with nucleic acids. These comprise transcriptional regulation, DNA replication and DNA repair. Conventional methods to study the interaction are either laborious (EMSA, ChIP) or limited to specific targets (EMSA, FP, ChIP).
A novel assay employs the Cy3 fluorophore which increases its fluorescence when being in proximity to proteins: Cy3-labeled oligonucleotides are immobilized on a microplate bottom and the basal fluorescence is measured on a microplate reader. If a protein is added to the microplate well that binds the oligonucleotide, the fluorescence of the fluorophore will increase. This increase can be measured and reports on protein-nucleic acid binding.
This simple and cost-effective assay detects sequence and structure specificities as well as binding constants of nucleic acid:protein liaisions. The change in fluorescence intensity is captured on a huge surface to provide stable measurements. This has been enabled by the well-scan function and high sensitivity of the FLUOstar® Omega.