Analysis of Nanomaterial Toxicity by Western Blot
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 wordsNanomaterials Electrophoresis SDS-PAGE Electroblotting Nitrocellulose PVDF Antibody Chemiluminescence
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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