Monitoring Protein Kinase Expression and Phosphorylation in Cell Lysates with Antibody Microarrays

  • Hong Zhang
  • Xiaoqing Shi
  • Steven Pelech
Part of the Methods in Molecular Biology book series (MIMB, volume 1360)


Fuelled by advances in our understanding of the human kinome and phosphoproteome and the increasing availability of pan- and phosphosite-specific antibodies, antibody microarrays have emerged as powerful tools for interrogating protein phosphorylation-mediated signaling systems in ex vivo studies. This economical platform permits ultra-sensitive, semiquantitative measurements of the levels of hundreds of protein kinases and their substrates along with their phosphorylation status simultaneously with minute amounts of specimens. Recent technological innovations in the design and fabrication of antibody microarrays and sample preparation have permitted further refinements of the technology to yield improvements in data quality. In this chapter, we describe a detailed protocol that we have developed for tracking the expression and phosphorylation of protein kinases and their substrates in crude cell lysate samples using a high-content antibody microarray.

Key words

Antibody microarray Phosphorylation Phosphoproteomics Protein kinase Protein kinome 


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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Kinexus Bioinformatics CorporationVancouverCanada
  2. 2.Department of MedicineUniversity of British ColumbiaVancouverCanada

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