Kinase Screening and Profiling by Hicham Zegzouti & Said A. Goueli

Author:Hicham Zegzouti & Said A. Goueli
Language: eng
Format: epub
Publisher: Springer New York, New York, NY

2 Materials

2.1 Cell line and Perturbation

1. A431 human epidermoid carcinoma cell line (CRL™-1555) (ATCC).

2.Recombinant Epidermal Growth Factor (EGF ) expressed in E. coli (Sigma-Aldrich).

2.2 KAM-850 Antibody Microarrays

The KAM-850 antibody microarray used in the protocols described here was developed and marketed by Kinexus Bioinformatics Corporation (Vancouver, BC, Canada, www.​kinexus.​ca) under the trademark of Kinex™ Antibody Microarray as a component used in its proteomics services or in a stand-alone assay kit. It features over 850 well-characterized polyclonal and monoclonal antibodies selected from more than 4600 antibodies either available commercially or developed by Kinexus based on their performance on more than 25 model systems of various species origins. Among them, over 510 pan-specific antibodies provide for the detection of 189 distinct protein kinases, 31 protein phosphatases, and 142 regulatory subunits of these enzymes and other cell signaling proteins that regulate cell proliferation, growth, stress response, and apoptosis. Approximately 340 phosphosite-specific antibodies track the non-redundant phosphorylation of 128 phosphosites in protein kinases, 4 phosphosites in protein phosphatases, and 155 phosphosites in other cell signaling proteins. In addition, antibodies for proteins encoded by some housekeeping genes such as actin and tubulin and those for nonspecific immunoglobulins from antibody host species such as rabbit, mouse, and goat are also included as controls.

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