Decrease time, increase throughput in prion and amyloid seeding assays
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چکیده
* Limit of detection < 100 amol of biotinylated and phosphorylated polypeptide (P-Tyr-100 assay kit, PerkinElmer, #6760620C), measured in white 384 small volume microplates (17 μL/well) Limit of detection was calculated according to the IUPAC standard: 3x(SDblank)/slope ©2016 All rights reserved. All logos and trademarks are the property of BMG LABTECH. Detection Modes Fluorescence Intensity including FRET UV/Vis Absorbance Spectra Luminescence (fl ash and glow) including BRET Time-Resolved Fluorescence including TR-FRET Alpha Technology Measurement Modes Top and bottom reading Endpoint and Kinetic measurements Sequential Multi-Excitation measurements Sequential Multi-Emission measurements Ratiometric measurements Well Scanning Microplate Formats Up to 384-well plates, 1536-well plates in absorbance, user-defi nable Light Source High energy xenon fl ash lamp Detectors Side window photomultiplier tube Optical Filters Excitation and emission fi lter wheels for 8 fi lters each Spectral Range 240 740 nm or 240 900 nm Absorbance Spectrometer: 220 1000 nm Sensitivity FI < 0.2 fmol/well Fluorescein TRF < 30 amol/well Europium High-End TRF for Omega < 3 amol/well Europium LUM 20 amol/well ATP DLReadyTM certifi ed
منابع مشابه
Rapid End-Point Quantitation of Prion Seeding Activity with Sensitivity Comparable to Bioassays
A major problem for the effective diagnosis and management of prion diseases is the lack of rapid high-throughput assays to measure low levels of prions. Such measurements have typically required prolonged bioassays in animals. Highly sensitive, but generally non-quantitative, prion detection methods have been developed based on prions' ability to seed the conversion of normally soluble proteas...
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Different transmissible spongiform encephalopathy (TSE)-associated forms of prion protein (e.g. PrP(Sc)) can vary markedly in ultrastructure and biochemical characteristics, but each is propagated in the host. PrP(Sc) propagation involves conversion from its normal isoform, PrP(C), by a seeded or templated polymerization mechanism. Such a mechanism is also the basis of the RT-QuIC and eQuIC pri...
متن کاملFibril Conformation as the Basis of Species- and Strain-Dependent Seeding Specificity of Mammalian Prion Amyloids
Spongiform encephalopathies are believed to be transmitted by self-perpetuating conformational conversion of the prion protein. It was shown recently that fundamental aspects of mammalian prion propagation can be reproduced in vitro in a seeded fibrillization of the recombinant prion protein variant Y145Stop (PrP23-144). Here we demonstrate that PrP23-144 amyloids from different species adopt d...
متن کاملFactors That Improve RT-QuIC Detection of Prion Seeding Activity
Rapid and sensitive detection of prions is important in managing prion diseases. The real-time quaking-induced conversion (RT-QuIC) assay for prion seeding activity has been applied to many prion diseases and provides for specific antemortem diagnostic testing. We evaluated RT-QuIC's long-term consistency and varied multiple reaction parameters. Repeated assays of a single scrapie sample using ...
متن کاملPrion-like seeding and nucleation of intracellular amyloid-β
Alzheimer's disease (AD) brain tissue can act as a seed to accelerate aggregation of amyloid-β (Aβ) into plaques in AD transgenic mice. Aβ seeds have been hypothesized to accelerate plaque formation in a prion-like manner of templated seeding and intercellular propagation. However, the structure(s) and location(s) of the Aβ seeds remain unknown. Moreover, in contrast to tau and α-synuclein, an ...
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تاریخ انتشار 2016