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OaAEP1-Mediated Enzymatic Synthesis and Immobilization of Polymerized Protein for Single-Molecule Force Spectroscopy
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JoVE Journal Bioquímica
OaAEP1-Mediated Enzymatic Synthesis and Immobilization of Polymerized Protein for Single-Molecule Force Spectroscopy

OaAEP1-Mediated Enzymatic Synthesis and Immobilization of Polymerized Protein for Single-Molecule Force Spectroscopy

DOI:
10.3791/60774-v

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08:34 min

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February 05, 2020

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, , , , , ,
1State Key Laboratory of Coordination Chemistry, School of Chemistry and Chemical Engineering,Nanjing University

Capítulos

  • 00:05Introduction
  • 00:43Functionalization of Coverslip and Cantilever Surface
  • 04:31Stepwise Polyprotein Preparation with Controlled Sequences
  • 05:44AFM Experiment Measurement and Data Analysis
  • 06:54Results: AFM-based SMFS Measurements, UV-Vis Absorbance Spectra and SDS-PAGE Gel Results
  • 07:51Conclusion

Summary

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Tadução automática

Here, we present a protocol to conjugate protein monomer by enzymes forming protein polymer with a controlled sequence and immobilize it on the surface for single-molecule force spectroscopy studies.

Tags

OaAEP1Enzymatic SynthesisPolymerized ProteinSingle-molecule Force SpectroscopyProtein OligomerPolymerization DegreeProtein-based MaterialCysteinePotassium ChromateSulfuric AcidAPTESSulfo-SMCCGL-ELP-50 Cys-proteinCantilever Surface

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