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Methods Mol Biol. 2018;1785:223-229. doi: 10.1007/978-1-4939-7841-0_14.

Antibody Specificity Profiling Using Protein Microarrays.

Author information

1
CDI Laboratories, Guanajibo Research and Innovation Park, Mayaguez, PR, USA.
2
Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD, USA. sblack@jhmi.edu.
3
Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD, USA. sblack@jhmi.edu.
4
Department of Ophthalmology, Johns Hopkins University School of Medicine, Baltimore, MD, USA. sblack@jhmi.edu.
5
Center for Human Systems Biology, Johns Hopkins University School of Medicine, Baltimore, MD, USA. sblack@jhmi.edu.
6
Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD, USA. sblack@jhmi.edu.

Abstract

Antibodies are the most widely used reagent for isolation and detection of specific proteins. However, using antibodies that are not highly specific in these studies can generate inaccurate and misleading data. Protein microarrays offer a platform by which antibody cross-reactivity against a broad range of cellular antigens can be simultaneously and quantitatively profiled. This protocol describes in detail the process of array pretreatment, antibody binding, washing, scanning and quantitative analysis of antibody specificity.

KEYWORDS:

Antibody specificity; Cross-reactivity; Monoclonal antibodies; Protein microarray

PMID:
29714021
DOI:
10.1007/978-1-4939-7841-0_14
[Indexed for MEDLINE]

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