Figure 1

Biotinylated LIMK peptide assay specificity. Serial dilutions of Phospho-LIMK1 (DRKKRYpTVVGNPY; squares) and LIMK1 (DRKKRYTVVGNPY; circles) peptide with concentrations ranging from 4 μg/ml to 90 ng/ml were bound to streptavidin plates (Pierce) for 2 hours at room temperature (24°C). Wells were then washed and probed with phospho-LIMK (Cell Signaling) over night at room temperature. Secondary decoration with IRDye®680 anti-rabbit(Li-Cor) was performed at room temperature for 1.5 hours. Proteins were visualized by direct fluorescence scanning with a Li-Cor Odyssey Imager. Error bars represent the standard deviation of triplicate determinations and are representative of three independent experiments.