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Nanotoxicity pp 161-169 | Cite as

Analysis of Nanomaterial Toxicity by Western Blot

  • Gao Long
  • Yiqun Mo
  • Qunwei Zhang
  • Mizu Jiang
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1894)

Abstract

Western blot is a routine biochemical technique for the immunodetection of proteins in cells and tissues exposed to nanomaterials (NMs). It is a sensitive method for protein analysis that involves the solubilization and separation of proteins by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE), transferring and immobilizing proteins onto a solid support, and targeted immunoprobing of a specific antigen. As a convenient and reliable research tool, the western blot plays an irreplaceable role in the era of proteomics together with mass spectrometry and protein chip. In this chapter we describe the detailed protocol for the entire process from sample preparation to quantitative measurement of target proteins.

Key words

Nanomaterials Electrophoresis SDS-PAGE Electroblotting Nitrocellulose PVDF Antibody Chemiluminescence 

Notes

Acknowledgment

This work was partly supported by the Public Welfare Project of Science Technology, Department of Zhejiang Province (2016C33152) to Dr. Mizu Jiang.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Gastroenterology, Affiliated Children’s HospitalZhejiang University School of MedicineHangzhouChina
  2. 2.Department of Environmental and Occupational Health Sciences, School of Public Health and Information SciencesUniversity of LouisvilleLouisvilleUSA

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