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Combinatorial Chemistry & High Throughput Screening


ISSN (Print): 1386-2073
ISSN (Online): 1875-5402

Development and Optimization of a Non-Radioactive JNK3 Assay

Author(s): Christian Peifer, Sabine Luik, Sabine Thuma, Yvonne Herweh and Stefan Laufer

Volume 9, Issue 8, 2006

Page: [613 - 618] Pages: 6

DOI: 10.2174/138620706778249721

Price: $65


In light of emerging interest in the relevance of c-Jun NH2-terminal protein kinase 3 (JNK3) as a promising drug target, we describe here an advanced non-radioactive immunosorbent JNK3 activity assay that is applicable for routine screening of small molecule ATP-competitive enzyme inhibitors. We modified and established a JNK3/ATF-2 protocol based on our previously described p38 MAPK method [1] for a substratebound non-radioactive procedure that represents a convenient alternative to conventional radioactive protein kinase assays. The objective of the present study was to validate these conditions by using the reference compounds SP600125 and SB203580 to achieve comparable IC50 results to published data. Furthermore, an IC50 for staurosporine was determined. The protocol we describe here represents an accessible and robust screening assay for JNK3 inhibitors.

Keywords: Non-radioactive immunosorbent JNK3 assay, staurosporine, SP600125, SB203580

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