ArticleNature protocols2012
Using bleach-chase to measure protein half-lives in living cells.
Article in Nature protocols, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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Who cites it
11 citing papers in PubMed, 19 citations in OpenAlex.
- Review
- The Metabolic Flux Probe (MFP)-Secreted Protein as a Non-Disruptive Information Carrier forInternational journal of molecular sciences · 2021Article
- Dynamics of protein synthesis and degradation through the cell cycle.Cell cycle (Georgetown, Tex.) · 2019Review
- MeCP2_E1 N-terminal modifications affect its degradation rate and are disrupted by the Ala2Val Rett mutation.Human molecular genetics · 2017Article
- Incompatibility of chemical protein synthesis inhibitors with accurate measurement of extended protein degradation rates.Pharmacology research & perspectives · 2017Article
- A Model of the Spatio-temporal Dynamics of Drosophila Eye Disc Development.PLoS computational biology · 2016Article
- Tracking the activity-dependent diffusion of synaptic proteins using restricted photoconversion of Dendra2.Frontiers in cellular neuroscience · 2015Article
- The emergence of proteome-wide technologies: systematic analysis of proteins comes of age.Nature reviews. Molecular cell biology · 2014Review
- Tracking protein turnover and degradation by microscopy: photo-switchable versus time-encoded fluorescent proteins.Open biology · 2014Article
- Noise genetics: inferring protein function by correlating phenotype with protein levels and localization in individual human cells.PLoS genetics · 2014Article
- Article
Corrections and comments
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Authors and funding
10 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Protein removal has a central role in numerous cellular processes. Obtaining systematic measurements of multiple protein removal rates is necessary to understand the principles that govern these processes, but it is currently a major technical challenge. To address this, we developed 'bleach-chase', a noninvasive method for measuring the half-lives of multiple proteins at high temporal resolution in living cells. The method uses a library of annotated human reporter cell clones, each with a unique fluorescently tagged protein expressed from its native chromosomal location. In this protocol, we detail a simple procedure that bleaches the cells and uses time-lapse fluorescence microscopy and automated image analysis to systematically measure the half-life dynamics of multiple proteins. The duration of the protocol is 4-5 d. The method may be applicable to a wide range of fluorescently tagged proteins and cell lines.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.