The amyloidophilic dyes thioflavin T and Congo red are small, yet powerful, molecules that allow the in vitro and in vivo detection of amyloid fibrils in protein solutions. Even though Congo red and thioflavin T binding assays are widespread techniques for unveiling amyloid fibers and are gradually replacing the more demanding X-ray diffraction method, handling samples containing amyloid fibrils is still challenging and can lead to false-positive/negative results. Here we describe a relatively straightforward procedure of preparing hen egg-white lysozyme amyloid fibrils in different buffer solutions and their detection with thioflavin T and Congo red, supported by an indispensable method for determining the secondary structure of proteins – circular dichroism.
CITATION STYLE
Brudar, S., & Hribar-Lee, B. (2023). The Role of Buffers in Wild-Type HEWL Amyloid Fibril Formation Mechanism: A Methodological Approach. In Methods in Molecular Biology (Vol. 2551, pp. 285–296). Humana Press Inc. https://doi.org/10.1007/978-1-0716-2597-2_19
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