Recombinant full-length human SLK was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. SLK or LOSK is a Ste20-like protein kinase that associates with microtubule and centrosome. Inhibition of SLK activity by dominant-negative mutant or RNAi leads to unfocused microtubule arrangement, indicating that SLK is needed for microtubule organization and proper localization of Golgi complex (1). Expression of v-Src can downregulate SLK activity through the involvement of CK2, which can directly phosphorylate and inhibit SLK activity, whereas inhibition of CK2 in v-Src-transformed cells results in normal SLK activity (2). CK2 and SLK can be co-localized in fibroblasts spreading on fibronectin-coated substrates.
ADP-Glo™ Kinase Assay is a luminescent kinase assay that measures ADP formed from a kinase reaction; ADP is converted into ATP, which is a substrate in a reaction catalyzed by Ultra-Glo™ Luciferase that produces light. The luminescent signal positively correlates with ADP amount and kinase activity. The assay is well suited for measuring the effects chemical compounds have on the activity of a broad range of purified kinases, making it ideal for both primary screening as well as kinase selectivity profiling. The ADP-Glo™ Kinase Assay can be used to monitor the activity of virtually any ADP-generating enzyme (e.g., kinase or ATPase) using up to 1mM ATP.
Profile More Compounds In-House: ADP-Glo™ Kinase Assay + Kinase Enzyme System is optimized so that you are up and running in no time. Complete Systems: The Kinase Enzyme Systems include a recombinant kinase enzyme, a substrate appropriate for the enzyme, a reaction buffer, DTT and supplemental reagents as needed. Obtain Reliable Results: The broad dynamic range, the ease of use and better sensitivity obtained with ADP-Glo™ Kinase Assay result in less ambiguous data.
Notes
Kinase Enzyme System manufactured by SignalChem.
Bulk quantities available upon request.
References
1.Burakov A.V. et al. (2005) LOSK (SLK) protein kinase activity is necessary for microtubule organization in the interphase cell centrosome. Dokl Biol Sci. 403, 317–9.
2.Chaar, Z. et al. (2006) v-Src-dependent down-regulation of the Ste20-like kinase SLK by casein kinase II. J Biol Chem. 281, 28193–9.