Measuring Amyloid‐β Peptide Concentrations in Murine Brain with Improved ELISA Assay
| Autor: | K. Sandy Pang, H Benson Peng, Inés A M de Lannoy |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
chemistry.chemical_classification
Genetically modified mouse 0303 health sciences General Immunology and Microbiology Chemistry Hydrochloride General Neuroscience Health Informatics Endogeny General Biochemistry Genetics and Molecular Biology 3. Good health Amino acid 03 medical and health sciences Medical Laboratory Technology chemistry.chemical_compound 0302 clinical medicine Cerebrospinal fluid Biochemistry Senile plaques General Pharmacology Toxicology and Pharmaceutics Guanidine 030217 neurology & neurosurgery 030304 developmental biology Homogenization (biology) |
| Zdroj: | Current Protocols. 1 |
| ISSN: | 2691-1299 |
| DOI: | 10.1002/cpz1.253 |
| Popis: | The amyloid-β (Aβ) peptides of 40 and 42 amino acids that are implicated in Alzheimer's disease may potentially aggregate into toxic oligomers and form neuritic plaques. The enzyme-linked immunosorbent assay (ELISA) is a facile method used for the determination of Aβ concentrations in biological matrices, namely plasma, cerebrospinal fluid, and brain. The method is mostly used for the measurement of Aβ concentrations in transgenic mice, but it is unknown whether the ELISA method is suitable for measuring low, endogenous levels of Aβ in the brains of wild-type mice. The Aβ ELISA kit manufacturer recommends use of 5 M guanidine hydrochloride (GuHCl), a protein-denaturing agent, for homogenization of the brain tissue, followed by dilution back down to 0.1 M to avoid quenching by GuHCl. Components of brain matrices and GuHCl that could interfere with the quantitation have not been investigated. In this article, we describe an improved method involving homogenization of mouse brain with 1 M instead of 5 M GuHCl, reducing the dilution factor by 5× to provide a higher sensitivity. The modified ELISA assay is improved for the quantitation of brain Aβ peptides in wild-type mice, where Aβ peptide levels are much lower than those in transgenic mouse models. © 2021 Wiley Periodicals LLC. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Plný text