Pan Proteome Labeling
Pan Proteome Labeling - Related Videos
11:53
Quantitative Proteomics Using Reductive Dimethylation for Stable Isotope Labeling
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Cited by 38 •
2014
Stable isotope labeling of peptides by reductive dimethylation (ReDi labeling) is a rapid, inexpensive strategy for accurate mass spectrometry-based quantitative proteomics. Here we demonstrate a robust method for preparation and analysis of protein mixtures using the ReDi approach that can be applied to nearly any sample type.
11:44
Metabolic Labeling and Membrane Fractionation for Comparative Proteomic Analysis of Arabidopsis thaliana Suspension Cell Cultures
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Cited by 5 •
2013
Here we describe a robust method for the fractionation of plant plasma membranes into detergent resistant and detergent soluble membranes based on a mixture of unlabeled and in vivo fully 15N labeled Arabidopsis thaliana cell cultures. The procedure is applied for comparative proteomic studies to understand signaling processes.
09:29
Skeletal Muscle Gender Dimorphism from Proteomics
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Cited by 4 •
2011
A straight-forward set of methods to isolate and determine the identity of the most abundant proteins expressed in skeletal muscle. About 800 spots are discerned on a two-dimensional gel from 10 mg muscle; this allows for the determination of gender-specific protein expression. These methods will give equivalent results in most tissues.
14:23
Bottom-up and Shotgun Proteomics to Identify a Comprehensive Cochlear Proteome
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Cited by 6 •
2014
Proteome analysis of the cochlear sensory epithelium can be challenging due to its small size and because membrane proteins are difficult to isolate and identify. Both membrane and soluble proteins can be identified by combining multiple preparative methods and separation techniques along with high-resolution mass spectrometry.
09:20
Proteomic Sample Preparation from Formalin Fixed and Paraffin Embedded Tissue
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Cited by 50 •
2013
Archival formalin fixed and paraffin embedded (FFPE) clinical samples are valuable material for investigation of diseases. Here we demonstrate a sample preparation workflow allowing in-depth proteomic analysis of microdissected FFPE tissue.
FAQs
During translation, cells incorporate chemically modified amino acids into proteins being produced at that time. These modifications provide a chemical handle on the newly formed proteins, allowing them to be distinguished from the broader protein pool after labeling. The approach therefore supports measurements of protein synthesis and turnover across many proteins rather than only one selected target.
The bioorthogonal reaction provides a way to attach a fluorescent or affinity tag to modified proteins after cellular incorporation has occurred. Because the reaction is designed to proceed without substantially disrupting cellular processes, researchers can label proteins while preserving the biological context being studied. The attached tag then makes the labeled proteins suitable for imaging, enrichment, or further analysis.
Single-target labeling concentrates on one selected protein, whereas pan proteome labeling captures information from proteins produced throughout the cellular proteome. This broader scope helps reveal coordinated changes in protein synthesis, turnover, localization, or abundance. As a result, researchers can examine global cellular responses instead of interpreting biology from one protein in isolation.
A typical workflow exposes cells to chemically modified amino acids so that newly produced proteins incorporate the modifications during translation. Researchers then use a bioorthogonal reaction to attach a fluorescent or affinity tag. The labeled material can subsequently be examined by mass spectrometry, visualized by fluorescence imaging, or enriched for additional protein analysis.
The analytical method depends on the information required. Mass spectrometry can assess broad protein-level changes, fluorescence imaging can show where labeled proteins are located in cells, and affinity enrichment can concentrate tagged proteins for downstream analysis. These readouts are complementary, allowing investigators to connect global protein changes with localization or selective recovery of labeled material.
This strategy can track how protein production and related protein dynamics change during cell growth, stress responses, development, disease mechanisms, or drug exposure. Because it surveys proteins across the proteome, it can reveal global changes associated with these conditions. Such measurements help connect cellular states or treatments with altered protein synthesis, turnover, localization, or abundance.