Applications of fluorescent polymer superquenching to high throughput screening assays for protein kinases.
Protein kinases are involved in the regulation of cellular metabolism, growth, differentiation, and proliferation. Aberrations in their function can lead to diseases such as cancer and inflammation. Protein kinases are therefore possible targets for drug therapies. To address the need for high throughput screening of potential inhibitors, QTL has developed a homogeneous and robust kinase assay for use in multiwell plate format. The QTL Lightspeed fluorescence superquenching-based kinase assays do not require specialized equipment, nor do they involve the use of radioactive hazardous materials or antibodies. QTL Lightspeed kinase assays directly measure the enzymatic activity of the target and do not involve secondary (detector) enzyme. In this article, we compare QTL Lightspeed protein kinase assays using Protein Kinase A, Protein Kinase Balpha/Akt1, and ribosomal S6 kinase-2 as examples with other commercially available kinase kits. Our data show that QTL Lightspeed kinase assays offer significant advantages over the current commercial kits in terms of both sensitivity and performance. The QTL Lightspeed kinase assay also offers a kinetic assay mode where the substrate phosphorylation can be monitored in real-time.
Xia
Wensheng
W
QTL Biosystems, LLC, Santa Fe, NM, USA.
Rininsland
Frauke
F
Wittenburg
Shannon K
SK
Shi
Xiaobo
X
Achyuthan
Komandoor E
KE
McBranch
Duncan W
DW
Whitten
David G
DG
D004305:Dose-Response Relationship, Drug; D011494:Protein Kinases / Q000032:analysis / Q000378:metabolism*; D013050:Spectrometry, Fluorescence / Q000379:methods; D013678:Technology, Pharmaceutical / Q000379:methods
D011494:Protein Kinases