Objective evaluation of differences in protein subcellular distribution
Related Experiment Video
Updated: Oct 2, 2026

Enriching Subcellular Proteins in Leptospira Using a Triton X-114-Based Fractionation Approach
Published on: August 8, 2025
Edward J S Roques1, Robert F Murphy
1Department of Biological Sciences and Center for Light Microscope Imaging and Biotechnology, Carnegie Mellon University, 4400 Fifth Avenue, Pittsburgh PA 15213, USA.
Abstract:
Research on the cell and molecular biology of proteins involved in membrane traffic often requires determination of the effects of various experimental conditions on the subcellular distributions of those proteins. This is most often accomplished by acquiring fluorescence microscope images and visually comparing these images. While this approach is quite suitable for detecting major changes in distributions, it is not sensitive to small changes and does not permit a quantitative and objective analysis. We therefore describe the application of pattern analysis methods to the comparison of sets of fluorescence microscope images. This approach provides a high throughput and reproducible technique to determine whether image distributions differ within a specified statistical confidence, and is shown to resolve image sets indistinguishable by visual inspection.
More Related Videos
Related Concept Videos
Subcellular Fractionation
Differential Centrifugation
Differential centrifugation is...
Protein Dynamics in Living Cells
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
Proteomics
Proteomics is the study of proteomes' function. It involves the large-scale systematic study of the proteome to denote the protein complement expressed by a genome. Scientist Mark Wilkins coined the term proteomics...
Overview of Protein Sorting and Transport
Protein sorting can be of two types: signal-based sorting and vesicle-based trafficking. In signal-based sorting, specific amino acid sequences called sorting signals target proteins to the proper location inside the cell either via gated transport or by protein translocation. In gated transport, folded...
Two-dimensional Gel Electrophoresis
The first dimension separation uses the isoelectric focusing or IEF technique performed on immobilized pH gradient (IPG) strips that separate proteins according to their isoelectric points.
Biological samples, such as cells...
Western Blotting
The technique begins with separating proteins from the sample using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), followed by protein transfer, immunoblotting, and finally, protein detection.

