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Single Cell Protein Analysis

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Cover of 'Single Cell Protein Analysis'

Table of Contents

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    Book Overview
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    Chapter 1 Single-Cell Western Blotting
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    Chapter 2 A Microfluidic Device for Immunoassay-Based Protein Analysis of Single E. coli Bacteria
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    Chapter 3 Enzyme-Linked ImmunoSpot (ELISpot) for Single-Cell Analysis
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    Chapter 4 Single Cell Protein Analysis
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    Chapter 5 Single Cell Protein Analysis
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    Chapter 6 Microfluidic Flow Cytometry for Single-Cell Protein Analysis
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    Chapter 7 Microfluidic Image Cytometry for Single-Cell Phenotyping of Human Pluripotent Stem Cells
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    Chapter 8 Characterizing Phenotypes and Signaling Networks of Single Human Cells by Mass Cytometry.
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    Chapter 9 Multiplexed Peptide-MHC Tetramer Staining with Mass Cytometry.
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    Chapter 10 Imaging and Mapping of Tissue Constituents at the Single-Cell Level Using MALDI MSI and Quantitative Laser Scanning Cytometry
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    Chapter 11 SPLIFF: A Single-Cell Method to Map Protein-Protein Interactions in Time and Space.
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    Chapter 12 Microfluidic Proximity Ligation Assay for Profiling Signaling Networks with Single-Cell Resolution
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    Chapter 13 Dynamics and Interactions of Individual Proteins in the Membrane of Single, Living Cells
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    Chapter 14 Microfluidics-Enabled Enzyme Activity Measurement in Single Cells
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    Chapter 15 Microfluidic Chemical Cytometry for Enzyme Assays of Single Cells
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    Chapter 16 Quantitative Detection of Nucleocytoplasmic Transport of Native Proteins in Single Cells
Attention for Chapter 16: Quantitative Detection of Nucleocytoplasmic Transport of Native Proteins in Single Cells
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Chapter title
Quantitative Detection of Nucleocytoplasmic Transport of Native Proteins in Single Cells
Chapter number 16
Book title
Single Cell Protein Analysis
Published in
Methods in molecular biology, January 2015
DOI 10.1007/978-1-4939-2987-0_16
Pubmed ID
Book ISBNs
978-1-4939-2986-3, 978-1-4939-2987-0
Authors

Zhenning Cao, Chang Lu

Abstract

The detection of protein translocation (i.e., the movement of intracellular proteins among various subcellular compartments) conventionally relies on imaging and subcellular-fractionation-based techniques that do not generate information on a large cell population with single-cell resolution. Although special flow cytometric tools such as imaging flow cytometry may generate single-cell data on processes such as nucleocytoplasmic transport, such equipment is expensive (thus has limited accessibility) and has low throughput for examining cells due to the reliance on high-speed imaging. Here we describe a protocol for detecting translocation of native proteins using a common flow cytometer which detects fluorescence intensity without imaging. We conduct chemical release of cytosolic proteins and fluorescence immunostaining of a targeted protein. The detected fluorescence intensity is quantitatively correlated to the cytosolic/nuclear localization of the protein at the single cell level. Our technique provides a simple route for studying nucleocytoplasmic transport with single-cell resolution using common flow cytometers.

Mendeley readers

The data shown below were compiled from readership statistics for 41 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 41 100%

Demographic breakdown

Readers by professional status Count As %
Researcher 3 7%
Other 1 2%
Unknown 37 90%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 2 5%
Environmental Science 1 2%
Medicine and Dentistry 1 2%
Unknown 37 90%