A Nonradioactive Fluorimetric SPE-Based Ceramide Kinase Assay Using NBD-C₆-Ceramide

Figure 6

Influence of proteine kinase inhibitors on CERK via 96 well HTS. CERK was measured with NBD-C₆-Cer in the presence of 0.5 mM of established protein kinase inhibitors (kinase inhibitor library, Biomol), using the 96 well fluorescent SPE assay. Activity is expressed as % of the control containing 10% DMSO (100 ± 11%; mean ± SD; 𝑛=3, magenta bars). CERK was not influenced by DMSO, up to 20% (data not shown). For comparison, effect of the inhibitors on sphingosine kinase is displayed in blue bars as % of control (100 ± 7%; mean ± SD; 𝑛=3).