Separation and detection of individual Aβ aggregates by capillary electrophoresis with laser-induced fluorescence detection.

Authors :: Picou RA
Schrum DP
Ku G
Cerqua RA
Kheterpal I
Gilman SD
Source :: Analytical biochemistry [Anal Biochem] 2012 Jun 15; Vol. 425 (2), pp. 104-12. Date of Electronic Publication: 2012 Mar 21.
Publication Year :: 2012
Abstract: The separation and detection of individual amyloid beta (Aβ) aggregates by capillary electrophoresis with laser-induced fluorescence detection (CE-LIF) was demonstrated. Samples were prepared with either Aβ (1-40) or Aβ (1-42) peptides and were characterized by CE with ultraviolet (UV) absorbance detection and transmission electron microscopy (TEM). Using thioflavin T (ThT) in the electrophoresis buffer, electrophoresis of aggregate-containing samples (5.0-s injection) produced up to several hundred narrow (< 20 ms FWHM [full width at half maximum]) fluorescence peaks. Injection of Aβ (1-40) monomer samples resulted in no additional peaks compared with controls. The CE-LIF results were validated by bulk ThT fluorescence measurements for the same samples. The potential of laser-induced fluorescence anisotropy (LIFA) with CE to characterize individual Aβ aggregates also was investigated.<br /> (Copyright © 2012 Elsevier Inc. All rights reserved.)

Subjects :: Amyloid beta-Peptides isolation & purification
Benzothiazoles
Microscopy, Electron, Scanning Transmission
Peptide Fragments isolation & purification
Thiazoles chemistry
Amyloid beta-Peptides analysis
Electrophoresis, Capillary
Lasers
Peptide Fragments analysis
Spectrophotometry, Ultraviolet

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