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Proteome-wide Quantification of Labeling Homogeneity at the Single Molecule Level

JoVE Journal
Biochemistry

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JoVE Journal Biochemistry

Proteome-wide Quantification of Labeling Homogeneity at the Single Molecule Level

Proteome-wide Quantification of Labeling Homogeneity at the Single Molecule Level

DOI:

10.3791/59199-v

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08:29 min

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April 19, 2019

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Simon Leclerc², Youri Arntz, Yuichi Taniguchi³

¹Laboratory for Cell Systems Control, RIKEN Center for Biosystems Dynamics Research,RIKEN, ²Laboratoire de Biomatériaux et Bioimagerie,INSERM, ³PRESTO, Japan Science and Technology Agency

Chapters

00:04Title
01:33Cell Lysis and Fluorescent Labeling
03:26Proteome Separation and Recovery
03:51Microscope Coverslip Preparation
05:46Observation with Single Molecule Fluorescence Microscopy
06:07Results: Labeling Homogeneity at the Single Molecule Level
07:33Conclusion

Summary

Automatic Translation

Here, we present a protocol to assess the labeling homogeneity for each protein species in a complex protein sample at the single molecule level.

Tags

Keywords: Protein Quantification Single-molecule Analysis Fluorescent Labeling Heterogeneity Molecular Concentration Assays Diagnostic Methods Protein Species Analysis Data Reproducibility Lysing Buffer Tween-20 DTT CHAPS Cy3 NHS Ester Dye HEPES-NaOH Biotin-2-amine EDC

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