Assays for light chain amyloidosis formation and cytotoxicity

Luis M. Blancas-Mejia, Pinaki Misra, Christopher J. Dick, Marta Marin-Argany, Keely R. Redhage, Shawna A. Cooper, Marina Ramirez-Alvarado

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Common biophysical techniques like absorption and fluorescence spectroscopy, microscopy, and light scattering studies have been in use to investigate fibril assembly for a long time. However, there is sometimes a lack of consensus from the findings of an individual technique when compared in parallel with the other techniques. In this chapter, we aim to provide a concise compilation of techniques that can effectively be used to obtain a comprehensive representation of the structural, aggregation, and toxicity determinants in immunoglobulin light chain amyloidosis. We start by giving a brief introduction on amyloid assembly and the advantages of using simple and readily available techniques to study aggregation. After an overview on preparation of protein to set up parallel experiments, we provide a systematic description of the in vitro techniques used to study aggregation in AL protein. Additionally, we thoroughly discuss the steps needed in our experience during the individual experiments for better reproducibility and data analysis.

LanguageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages123-153
Number of pages31
DOIs
StatePublished - Jan 1 2019

Publication series

NameMethods in Molecular Biology
Volume1873
ISSN (Print)1064-3745

Fingerprint

Amyloidosis
Immunoglobulin Light Chains
Light
Fluorescence Spectrometry
Fluorescence Microscopy
Amyloid
Proteins
In Vitro Techniques

Keywords

  • Amyloid formation
  • Amyloidosis
  • Caspase assay
  • Cell viability
  • Electron microscopy
  • Immunoglobulin light chain
  • Sedimentation assay
  • Thioflavin T

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Blancas-Mejia, L. M., Misra, P., Dick, C. J., Marin-Argany, M., Redhage, K. R., Cooper, S. A., & Ramirez-Alvarado, M. (2019). Assays for light chain amyloidosis formation and cytotoxicity. In Methods in Molecular Biology (pp. 123-153). (Methods in Molecular Biology; Vol. 1873). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-8820-4_8

Assays for light chain amyloidosis formation and cytotoxicity. / Blancas-Mejia, Luis M.; Misra, Pinaki; Dick, Christopher J.; Marin-Argany, Marta; Redhage, Keely R.; Cooper, Shawna A.; Ramirez-Alvarado, Marina.

Methods in Molecular Biology. Humana Press Inc., 2019. p. 123-153 (Methods in Molecular Biology; Vol. 1873).

Research output: Chapter in Book/Report/Conference proceedingChapter

Blancas-Mejia, LM, Misra, P, Dick, CJ, Marin-Argany, M, Redhage, KR, Cooper, SA & Ramirez-Alvarado, M 2019, Assays for light chain amyloidosis formation and cytotoxicity. in Methods in Molecular Biology. Methods in Molecular Biology, vol. 1873, Humana Press Inc., pp. 123-153. https://doi.org/10.1007/978-1-4939-8820-4_8
Blancas-Mejia LM, Misra P, Dick CJ, Marin-Argany M, Redhage KR, Cooper SA et al. Assays for light chain amyloidosis formation and cytotoxicity. In Methods in Molecular Biology. Humana Press Inc. 2019. p. 123-153. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-8820-4_8
Blancas-Mejia, Luis M. ; Misra, Pinaki ; Dick, Christopher J. ; Marin-Argany, Marta ; Redhage, Keely R. ; Cooper, Shawna A. ; Ramirez-Alvarado, Marina. / Assays for light chain amyloidosis formation and cytotoxicity. Methods in Molecular Biology. Humana Press Inc., 2019. pp. 123-153 (Methods in Molecular Biology).
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