Abstract
Antibody arrays represent one of the very early protein array systems where antibodies are used to capture and detect target proteins in a high-throughput platform. The development of high-quality antibodies, nanomaterial-based novel detection probes, as well as innovative imaging technologies and computational tools has tremendously improved the sensitivity, specificity, and robustness of antibody arrays during the past decade. In this protocol we will incorporate the most updated innovations and developments of antibody arrays into the step-by-step experimental procedures. This includes antibody printing, sample preparation, array detection, as well as imaging and data analysis. Antibody array could be used for cytokine profiling or mapping of phosphorylation, glycosylation, or other post-translational modifications of target proteins.
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
Orchekowski R, Hamelinck D, Li L, Gliwa E, VanBrocklin M, Marrero JA, Woude GFV, Feng Z, Brand R, Haab BB (2005) Antibody microarray profiling reveals individual and combined serum proteins associated with pancreatic cancer. Cancer Res 65(23):11193–11202
Borrebaeck CA, Wingren C (2014) Antibody array generation and use. Methods Mol Biol 1131:563–571
Mueller C, Liotta LA, Espina V (2010) Reverse phase protein microarrays advance to use in clinical trials. Mol Oncol 4(6):461–481
Wu T, Ding H, Han J, Arriens C, Wei C, Han W, Pedroza C, Jiang S, Anolik J, Petri M, Sanz I, Saxena R, Mohan C (2016) Antibody-array-based proteomic screening of serum markers in systemic lupus erythematosus: a discovery study. J Proteome Res 15(7):2102–2114. doi:10.1021/acs.jproteome.5b00905
Wu T, Du Y, Han J, Singh S, Xie C, Guo Y, Zhou XJ, Ahn C, Saxena R, Mohan C (2013) Urinary angiostatin—a novel putative marker of renal pathology chronicity in lupus nephritis. Mol Cell Proteomics 12(5):1170–1179. doi:10.1074/mcp.M112.021667
Zong Y, Zhang S, Chen H-T, Zong Y, Shi Y (2007) Forward-phase and reverse-phase protein microarray. Methods Mol Biol 381:363–374
Acknowledgments
This work was partly supported by a grant from the Lupus Research Institute to T.W. and a startup fund from the University of Houston to T.W.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Yuan, Y., Lin, ZT., Wang, H., Hong, X., Heon, M., Wu, T. (2017). Protein Arrays I: Antibody Arrays. In: Kaufmann, M., Klinger, C., Savelsbergh, A. (eds) Functional Genomics. Methods in Molecular Biology, vol 1654. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7231-9_19
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7231-9_19
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7230-2
Online ISBN: 978-1-4939-7231-9
eBook Packages: Springer Protocols