Nanotoxicity pp 161-169 | Cite as

Analysis of Nanomaterial Toxicity by Western Blot

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


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 



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


  1. 1.
    Ema M, Gamo M, Honda K (2016) A review of toxicity studies of single-walled carbon nanotubes in laboratory animals. Regul Toxicol Pharmacol 74:42–63CrossRefGoogle Scholar
  2. 2.
    Gliga AR, Skoglund S, Wallinder IO, Fadeel B, Karlsson HL (2014) Size-dependent cytotoxicity of silver nanoparticles in human lung cells: the role of cellular uptake, agglomeration and Ag release. Part Fibre Toxicol 11:11CrossRefGoogle Scholar
  3. 3.
    Oberdörster G, Oberdörster E, Oberdörster J (2005) Nanotoxicology: an emerging discipline evolving from studies of ultrafine particles. Environ Health Perspect 113:823–839CrossRefGoogle Scholar
  4. 4.
    Mahmood M, Casciano DA, Mocan T, Iancu C, Xu Y, Mocan L, Iancu DT, Dervishi E, Li Z, Abdalmuhsen M, Biris AR, Ali N, Howard P, Biris AS (2010) Cytotoxicity and biological effects of functional nanomaterials delivered to various cell lines. J Appl Toxicol 30:74–83CrossRefGoogle Scholar
  5. 5.
    Poon WL, Alenius H, Ndika J, Fortino V, Kolhinen V, Meščeriakovas A, Wang M, Greco D, Lähde A, Jokiniemi J, Lee JC, El-Nezami H, Karisola P (2017) Nano-sized zinc oxide and silver, but not titanium dioxide, induce innate and adaptive immunity and antiviral response in differentiated THP-1 cells. Nanotoxicology 11:936–951CrossRefGoogle Scholar
  6. 6.
    Ivask A, Juganson K, Bondarenko O, Mortimer M, Aruoja V, Kasemets K, Blinova I, Heinlaan M, Slaveykova V, Kahru A (2014) Mechanisms of toxic action of Ag, ZnO and CuO nanoparticles to selected ecotoxicological test organisms and mammalian cells in vitro: a comparative review. Nanotoxicology 8 Suppl 1:57–71CrossRefGoogle Scholar
  7. 7.
    Tuli HS, Kashyap D, Bedi SK, Kumar P, Kumar G, Sandhu SS (2015) Molecular aspects of metal oxide nanoparticle (MO-NPs) mediated pharmacological effects. Life Sci 143:71–79CrossRefGoogle Scholar
  8. 8.
    Chevallet M, Veronesi G, Fuchs A, Mintz E, Michaud-Soret I, Deniaud A (2017) Impact of labile metal nanoparticles on cellular homeostasis. Biochim Biophys Acta 1861:1566–1577CrossRefGoogle Scholar
  9. 9.
    Hendren CO, Lowry GV, Unrine JM, Wiesner MR (2015) A functional assay-based strategy for nanomaterial risk forecasting. Sci Total Environ 536:1029–1037CrossRefGoogle Scholar

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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