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Assay Method Information |
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| | Fluorescence (or Forster) Resonance Energy Transfer (FRET) Assay |
| Description: | In this assay, assembly of Abeta 1-42 oligomer formation is monitored by FRET using N-terminal conjugates of fluorescein-Abeta 1-42 as both the donor and acceptor fluorophore (fluorescein-fluorescein Ro ~45 A). Abeta 1-42 monomers assembling into oligomeric species results in a decrease of fluorescence as the fluorescein labeled Abeta 1-42 peptides become proximal to each other and FRET efficiency increases. Inhibition of Abeta 1-42 assembly is observed as the absence or attenuation of fluorescein quenching.FRET and FP assays are performed in 384-well Corning Non-Binding Surface, black, opaque microtiter plates, and the assay buffer consists of 50 mM MOPS-Tris (pH 8.0) with 100 mM MgCl2. The assay volume, containing 0.2 uM FITC-Abeta (1-42) and 0.8 uM Abeta (1-42), is 50 ul and the temperature is 37 C. ADDL assembly is monitored on a Tecan GENios Pro plate reader, exciting at a wavelength of 485 nm and detecting emission at a wavelength of 515 nm. |
| Affinity data for this assay | |
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