Filter trapping protocol to detect aggregated proteins in human cell lines

Jagat K Chhipi-Shrestha; Minoru Yoshida; Shintaro Iwasaki

doi:10.1016/j.xpro.2022.101571

Filter trapping protocol to detect aggregated proteins in human cell lines

STAR Protoc. 2022 Jul 19;3(3):101571. doi: 10.1016/j.xpro.2022.101571. eCollection 2022 Sep 16.

Authors

Jagat K Chhipi-Shrestha¹, Minoru Yoshida², Shintaro Iwasaki³

Affiliations

¹ Chemical Genomics Research Group, RIKEN Center for Sustainable Resource Science, Wako, Saitama 351-0198, Japan.
² Chemical 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. Electronic address: yoshidam@riken.jp.
³ RNA 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. Electronic address: shintaro.iwasaki@riken.jp.

Abstract

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).

Keywords: Molecular Biology; Protein Biochemistry; Protein expression and purification.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Biological Assay
Cell Line
Humans
Protein Aggregates*
Proteins*
Staining and Labeling

Substances

Protein Aggregates
Proteins