Abstract
Electrophoresis and the following western blot analysis are indispensable to investigate biochemical changes in cells and tissues exposed to nanoparticles or nanomaterials. Proteins should be extracted from the cells and tissues using a proper method, especially when phosphorylated proteins are to be detected. It is important to select a good blocking agent and an appropriate pair of primary and peroxidase-tagged secondary antibodies to obtain good results in western blot analysis. One thing that may be specific to nanomaterials, and that you should keep in mind, is that some proteins may be adsorbed on the surface of particulate nanomaterials. In this chapter the whole process of western blot analysis, from sample preparation to quantitative measurement of target proteins, is described.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Fenoglio I, Croce A, Di Renzo F, Tiozzo R, Fubini B (2000) Pure-silica zeolites (Porosils) as model solids for the evaluation of the physicochemical features determining silica toxicity to macrophages. Chem Res Toxicol 13:489–500
Oberdorster G, Oberdorster E, Oberdorster J (2005) Nanotoxicology: an emerging discipline evolving from studies of ultrafine particles. Environ Health Perspect 113:823–839
Tabet L, Bussy C, Amara N, Setyan A, Grodet A, Rossi MJ, Pairon JC, Boczkowski J, Lanone S (2009) Adverse effects of industrial multiwalled carbon nanotubes on human pulmonary cells. J Toxicol Environ Health A 72:60–73
Wick P, Manser P, Limbach LK, Dettlaff-Weglikowska U, Krumeich F, Roth S, Stark WJ, Bruinink A (2007) The degree and kind of agglomeration affect carbon nanotube cytotoxicity. Toxicol Lett 168:121–131
O’Farrell PH (1975) High resolution two-dimensional electrophoresis of proteins. J Biol Chem 250:4007–4021
Hirano S, Fujitani Y, Furuyama A, Kanno S (2010) Uptake and cytotoxic effects of multi-walled carbon nanotubes in human bronchial epithelial cells. Toxicol Appl Pharmacol 249:8–15
Hirano S, Kanno S, Furuyama A (2008) Multi-walled carbon nanotubes injure the plasma membrane of macrophages. Toxicol Appl Pharmacol 232:244–251
Jiang X, Weise S, Hafner M, Rocker C, Zhang F, Parak WJ, Nienhaus GU (2010) Quantitative analysis of the protein corona on FePt nanoparticles formed by transferrin binding. J R Soc Interf 7:S5–S13
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Hirano, S. (2012). Western Blot Analysis. In: Reineke, J. (eds) Nanotoxicity. Methods in Molecular Biology, vol 926. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-002-1_6
Download citation
DOI: https://doi.org/10.1007/978-1-62703-002-1_6
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-001-4
Online ISBN: 978-1-62703-002-1
eBook Packages: Springer Protocols