Filter trapping protocol to detect aggregated proteins in human cell lines
Summary: The loss of protein homeostasis results in cytotoxic protein aggregates, a common hallmark of aging and neurological diseases. Here, we present an adjusted filter-trapping assay protocol to detect global aggregated proteins in human cell lines, via a high-sensitive protein staining method....
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2022-09-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166722004518 |
| _version_ | 1818193503556993024 |
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| author | Jagat K. Chhipi-Shrestha Minoru Yoshida Shintaro Iwasaki |
| author_facet | Jagat K. Chhipi-Shrestha Minoru Yoshida Shintaro Iwasaki |
| author_sort | Jagat K. Chhipi-Shrestha |
| collection | DOAJ |
| description | Summary: The loss of protein homeostasis results in cytotoxic protein aggregates, a common hallmark of aging and neurological diseases. Here, we present an adjusted filter-trapping assay protocol to detect global aggregated proteins in human cell lines, via a high-sensitive protein staining method. This protocol also details an alternative approach to monitor specific protein aggregates trapped in the filter membrane, by subsequent immunoblotting of ectopically expressed and endogenous proteins.For complete details on the use and execution of this protocol, please refer to Chhipi-Shrestha et al. (2022). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-12-12T00:47:26Z |
| format | Article |
| id | doaj.art-a2b2cde2e8eb41b681f7cfc9131fd657 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-12-12T00:47:26Z |
| publishDate | 2022-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-a2b2cde2e8eb41b681f7cfc9131fd6572022-12-22T00:44:06ZengElsevierSTAR Protocols2666-16672022-09-0133101571Filter trapping protocol to detect aggregated proteins in human cell linesJagat K. Chhipi-Shrestha0Minoru Yoshida1Shintaro Iwasaki2Chemical Genomics Research Group, RIKEN Center for Sustainable Resource Science, Wako, Saitama 351-0198, JapanChemical Genomics Research Group, RIKEN Center for Sustainable Resource Science, Wako, Saitama 351-0198, Japan; Department of Biotechnology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Bunkyo-ku, Tokyo 113-8657, Japan; Collaborative Research Institute for Innovative Microbiology, The University of Tokyo, Bunkyo-ku, Tokyo 113-8657, Japan; Corresponding authorRNA Systems Biochemistry Laboratory, RIKEN Cluster for Pioneering Research, Wako, Saitama 351-0198, Japan; Department of Computational Biology and Medical Sciences, Graduate School of Frontier Sciences, The University of Tokyo, Kashiwa, Chiba 277-8561, Japan; Corresponding authorSummary: The loss of protein homeostasis results in cytotoxic protein aggregates, a common hallmark of aging and neurological diseases. Here, we present an adjusted filter-trapping assay protocol to detect global aggregated proteins in human cell lines, via a high-sensitive protein staining method. This protocol also details an alternative approach to monitor specific protein aggregates trapped in the filter membrane, by subsequent immunoblotting of ectopically expressed and endogenous proteins.For complete details on the use and execution of this protocol, please refer to Chhipi-Shrestha et al. (2022). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166722004518Molecular BiologyProtein BiochemistryProtein expression and purification |
| spellingShingle | Jagat K. Chhipi-Shrestha Minoru Yoshida Shintaro Iwasaki Filter trapping protocol to detect aggregated proteins in human cell lines STAR Protocols Molecular Biology Protein Biochemistry Protein expression and purification |
| title | Filter trapping protocol to detect aggregated proteins in human cell lines |
| title_full | Filter trapping protocol to detect aggregated proteins in human cell lines |
| title_fullStr | Filter trapping protocol to detect aggregated proteins in human cell lines |
| title_full_unstemmed | Filter trapping protocol to detect aggregated proteins in human cell lines |
| title_short | Filter trapping protocol to detect aggregated proteins in human cell lines |
| title_sort | filter trapping protocol to detect aggregated proteins in human cell lines |
| topic | Molecular Biology Protein Biochemistry Protein expression and purification |
| url | http://www.sciencedirect.com/science/article/pii/S2666166722004518 |
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