Phosphorylation state and protein levels measured in BRAF(V600E/D) melanoma cell lines monitored by Reverse Phase Protein Arrays (RPPA)

This assay measures phosphorylation state and protein levels in BRAF(V600E/D) melanoma cell lines monitored by Reverse Phase Protein Arrays (RPPA). Cells were plated in 4 replicates at 20,000 (or 15,000) cells per well density in 96-well plates, treated with different doses of each compound for 1, 5, 10, 24 and 48 hr. Plates for RPPA assays were treated with drugs using previously prepared 384-well dilution plates and a Seiko pin transfer robot system. Lysates were collected at the designated time-points (1, 5, 10, 24, 48 hr) post drug treatment. To generate reverse phase arrays, lysates were printed on nitrocellulose coated glass slides on a 2470 Arrayer. The data from this dataset and from the related HMS dataset #20217 were used to generate the signatures presented in HMS datasets #20229-20231.