Quantifying Amyloid β-Peptide (Aβ) Aggregation Using the Congo Red-Aβ (CR–Aβ) Spectrophotometric Assay
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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Congo red binds amyloid-like fibrils with a β-sheet structure in a regular parallel arrangement, detectable by a red-shift in the Congo red absorbance spectrum.
“Congo red is known to bind amyloid-like fibrils with a β-sheet structure in a regular parallel arrangement, which can be detected by a red-shift in the Congo red absorbance spectrum (42).”
From Reelin molecules assemble together to form a large protein complex, which is inhibited by the function-blocking CR-50 antibody (2000) · cites this paper as [42]
The point of maximal spectral difference at 528 nm is shorter than that of amyloid fibrils (541 nm).
“which is shorter than that of amyloid fibrils (541 nm; ref. 42).”
From Reelin molecules assemble together to form a large protein complex, which is inhibited by the function-blocking CR-50 antibody (2000) · cites this paper as [42]
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- Authors
- Klunk, W. E., Jacob, R. F., & Mason, R. P.
- Affiliations
- University of Pittsburgh (US)
- ORCID
- Klunk, William E.
- 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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