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Updated: Mar 15, 2026
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Highly Sensitive and Quantitative Detection of Proteins and Their Isoforms by Capillary Isoelectric Focusing Method
Published on: September 19, 2018
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Detection of Isoforms Differing by a Single Charge Unit in Individual Cells
Augusto M Tentori1,2, Kevin A Yamauchi1, Amy E Herr3,4
1The UC Berkeley/UCSF Graduate Program in Bioengineering, Berkeley, CA, USA.
Angewandte Chemie (International Ed. in English)
|September 7, 2016
Summary
We developed single-cell isoelectric focusing (scIEF) to measure protein isoforms in individual mammalian cells. This method offers high-resolution analysis of cytoplasmic and nuclear proteins from single cells.
Area of Science:
- Biochemistry
- Cell Biology
- Analytical Chemistry
Background:
- Measuring protein isoforms in individual cells is challenging.
- Existing methods lack the resolution for single-cell analysis of protein variants.
Purpose of the Study:
- To develop a novel method for high-resolution protein isoform analysis at the single-cell level.
- To enable the study of protein heterogeneity within individual mammalian cells.
Main Methods:
- Developed single-cell isoelectric focusing (scIEF) using microfluidic technology.
- Utilized photoactivatable materials for tunable pH gradients and precise sample handling.
- Employed high-selectivity immunoprobing for protein detection.
Main Results:
- Achieved single-cell resolution for isoelectric focusing (scIEF).
- Resolved protein isoforms with single-charge unit resolution.
- Successfully analyzed endogenous cytoplasmic and nuclear proteins from individual mammalian cells.
Conclusions:
- scIEF is a powerful technique for analyzing protein isoform heterogeneity in single cells.
- This method opens new avenues for understanding cellular protein expression and function.
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