Cited by 311Alzheimer's disease research and treatmentsDementia and Cognitive Impairment ResearchProtein purification and stability

Quantifying Amyloid β-Peptide (Aβ) Aggregation Using the Congo Red-Aβ (CR–Aβ) Spectrophotometric Assay

Klunk, William E., Jacob, Robert F., Mason, R.Preston·1999·Analytical Biochemistry

Abstract

Congo red (CR) is a histologic dye that binds to many amyloid proteins because of their extensive beta-sheet structure. The absorbance spectrum of the dye changes upon binding to amyloid. This spectral change has previously been exploited to develop a method to study the interaction of CR with fibrillar beta-sheet insulin fibrils, a model amyloid protein. The amyloid beta-peptide (Abeta) is an amyloid protein which is deposited in the brains of Alzheimer's disease victims. Abeta is toxic to neurons in vitro in a manner that is highly dependent on the assembly of this peptide into beta-sheet fibrils. The CR-insulin assay has been applied as a means of studying the aggregation of Abeta, despite the fact that the CR-insulin procedure was never adequately developed for this purpose. In this study, we modify our original CR-insulin assay specifically for the purpose of quantifying Abeta aggregation and discuss the reasons why application of the CR-insulin method is not valid for this purpose. The CR-Abeta method is equally simple and retains the advantages of speed and lack of necessity for specialized instrumentation or expensive/radioactive reagents. Furthermore, this method can directly provide quantitation of aggregated Abeta in absolute terms (i.e., microg/ml).

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Crossref · 10.1006/abio.1998.2933

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Authors
Klunk, W. E., Jacob, R. F., & Mason, R. P.
Affiliations
University of Pittsburgh (US)
Venue
Analytical Biochemistry
Publisher
Elsevier BV
Published
1999-01
Volume
266
Issue
1
Pages
66-76
ISSN
0003-2697 (ISSN-L)
OpenAlex
W2053546096
Cited by
310
References
30
Citations by year
2026: 4 · 2025: 8 · 2024: 10 · 2023: 6 · 2022: 5 · 2021: 9 · 2020: 15 · 2019: 14 · 2018: 18 · 2017: 15 · 2016: 15 · 2015: 11 · 2014: 8 · 2013: 13 · 2012: 20
FWCI
6.42
Type
journal article
Language
en
Open access
closed
Locations
  • Analytical Biochemistry (publishedVersion)
  • PubMed (publishedVersion)
Funders
National Institute on Aging
Topics
  • Health Sciences › Medicine › Physiology › Alzheimer's disease research and treatments
  • Health Sciences › Medicine › Psychiatry and Mental health › Dementia and Cognitive Impairment Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein purification and stability
Keywords
amyloid-beta, Congo red, spectrophotometric assay, β-sheet fibrils, Alzheimer's disease, dye-protein binding, amyloidosis
MeSH terms
Congo Red, Coloring Agents, Spectrophotometry, Amyloid beta-Peptides

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