Related Experiment Video
Updated: Feb 2, 2026
09:45
Detection of Copy Number Alterations Using Single Cell Sequencing
Published on: February 17, 2017
12.2K
Single-Cell High-Resolution Detection and Quantification of Protein Isoforms Differing by a Single Charge Unit
Kristi A Koelsch1,2,3
1University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA. Kristi-Koelsch@omrf.org.
Methods in Molecular Biology (Clifton, N.J.)
|November 15, 2018
Summary
A new microfluidic method enables single-cell proteomics using isoelectric focusing (IEF). This technique allows for the analysis of heterogeneous cell populations by separating proteins based on their isoelectric points.
Area of Science:
- Biochemistry
- Proteomics
- Analytical Chemistry
Background:
- Isoelectric focusing (IEF) separates proteins by isoelectric point.
- Single-cell analysis is crucial for understanding cell heterogeneity.
- Traditional IEF was not suitable for single-cell applications.
Purpose of the Study:
- To present a novel method for single-cell proteomics.
- To enable the analysis of protein expression in individual cells.
Main Methods:
- Microfluidics coupled with isoelectric focusing (IEF).
- Photocapture and immunoprobing within a micro-gel.
- Multi-reprobing capability of the micro-gel.
Main Results:
- Successful application of IEF for single-cell proteomic analysis.
- Demonstration of protein separation and detection in single cells.
- Validation of the micro-gel's reusability for multiple analyses.
Conclusions:
- The developed method overcomes limitations of traditional IEF for single-cell studies.
- This technique facilitates deeper insights into cellular heterogeneity.
- The platform offers a versatile tool for advanced proteomic research.
Related Concept Videos
01:25Single-pass Transmembrane Proteins
6.6K
Integral membrane proteins are tightly associated with the cell membrane and play a crucial role in cell communication, signaling, adhesion, and transport of the molecules. Some integral membrane proteins are present only in the membrane monolayer. For example, the enzyme fatty acid amide hydrolase is present in the cytoplasmic side of the membrane monolayer. In contrast, another type of integral membrane protein, also known as a transmembrane protein, spans across the membrane. Transmembrane...
6.6K
01:05Difference from Background: Limit of Detection
8.3K
The limit of detection (LOD) is the smallest amount of analyte that can be distinguished from the background noise. The LOD value corresponds to the concentration at which the analyte signal is three times larger than the standard deviation of the blank signal. Below this value, the analyte signal cannot be differentiated from the background noise. It is calculated by dividing the calibration slope by 3 times the standard deviation of the blank signals.
The LOD indicates the presence or absence...
The LOD indicates the presence or absence...
8.3K
01:03Single-Strand DNA Binding Proteins
16.7K
For successful DNA replication, the unwinding of double-stranded DNA must be accompanied by stabilization and protection of the separated single strands of the DNA. This crucial task is performed by single-strand DNA-binding (SSB) proteins. They bind to the DNA in a sequence-independent manner, which means that the nitrogenous bases of the DNA need not be present in a specific order for binding of SSB proteins to it. The binding of SSB proteins straightens single-stranded DNA (ssDNA) and makes...
16.7K
01:26Insertion of Single-pass Transmembrane Proteins in the RER
18.1K
Integral membrane proteins are proteins adhered to the lipid bilayer of a cell organelle or membrane. They can be of two types: transmembrane integral proteins that span the lipid bilayer and monotopic proteins that are attached to either side of the membrane but do not pass through it.
Integral transmembrane proteins possess transmembrane and extra membrane domains. The transmembrane domains are primarily made of 20-25 hydrophobic amino acids arranged in a helical secondary confirmation. These...
Integral transmembrane proteins possess transmembrane and extra membrane domains. The transmembrane domains are primarily made of 20-25 hydrophobic amino acids arranged in a helical secondary confirmation. These...
18.1K
02:42Formal Charges
40.5K
In some cases, there are seemingly more than one valid Lewis structures for molecules and polyatomic ions. The concept of formal charges can be used to help predict the most appropriate Lewis structure when more than one reasonable structure exists.
40.5K
02:21Cancers Originate from Somatic Mutations in a Single Cell
14.9K
Cancer arises from mutations in the critical genes that allow healthy cells to escape cell cycle regulation and acquire the ability to proliferate indefinitely. Though originating from a single mutation event in one of the originator cells, cancer progresses when the mutant cell lines continue to gain more and more mutations, and finally, become malignant. For example, chronic myelogenous leukemia (CML) develops initially as a non-lethal increase in white blood cells, which progressively...
14.9K