Implement HTRF® phospho-protein assays at every step of the drug discovery process: from in vitro to in vivo models

J. Vallaghé1, D. Monchiet1, N. Costy1, M. Linnebacher2, T. Chardès3, C. Larbouret3, J. Pannequin4, S. Dalle4, E. Trinquet1, F. Charrier-Savournin1


1Cisbio Bioassays, Codolet, France | 2University of Rostock, Germany | 3Institut de Recherche en Cancérologie de Montpellier, INSERM U896, Université Montpellier 1, Institut régional du Cancer Montpellier / Val d’Aurelle, France | 4Institut de Génomique F

SLAS 2015

High-throughput screening (HTS) on two-dimensional (2D) immortalized cell cultures is frequently the starting point for identifying promising new drugs. To easily and rapidly translate drug discovery research from simplistic in vitro to physiologically relevant in vivo models, researchers need robust biochemical assays compatible with samples presenting different levels of complexity.
This study demonstrates that the HTRF phospho-/total protein platform is suitable for the rapid and robust analysis of cell signaling pathways in different kinds of biological models such as 2D, 3D  cell cultures, patient-derived primary cells, and xenografts.

Phosphorylated proteins

Phospho-4EBP1 (Thr37/46) Cellular Assay KitPhospho-ACC (Ser79) Cellular Assay KitPhospho-AMPK (Thr172) & Total AMPK Cellular Assay Kits Phospho-ATF2 (Thr71) Cellular Assay KitPhospho-β-Catenin (Thr41/Ser37/Ser33) & Total β-Catenin Cellular Assay Kits Phospho-Bad (Ser112) Cellular Assay Kit Phospho-c-Jun (Ser63) Cellular Assay KitPhospho-c-RAF (Ser338) Cellular Assay KitPhospho-EGFR (pan), Phospho-EGFR (Tyr1068) & Total EGFR Cellular Assay KitsPhospho-EIF4E (Ser209) Cellular Assay Kit Phospho-GSK3β (Ser9), Phospho-GSK3α (Ser21), Total GSK3β & Total GSK3α Cellular Assay KitsPhospho-H2AX (Ser139) Cellular Assay KitPhospho-HER2 (Tyr1221/1222) & Total HER2 Cellular Assay KitsPhospho-HER3 (Tyr1289) & Total HER3 Cellular Assay KitsPhospho-Histone H3 (Ser10), Phospho-Histone H3 (Thr3) & Total Histone H3 Cellular Assay KitsPhospho-IKKβ (Ser177/181) & Total IKKβ Cellular Assay Kit Phospho-JNK (Thr183 / Tyr185) Cellular Assay Kit Phospho-MEK1 (Ser218/222), Phospho-MEK1/2 (Ser218/222), Phospho-MEK1 (Ser298) & Total MEK1 Cellular Assay KitsPhospho-mTOR (Ser2448) Cellular Assay KitPhospho-NFkB (Ser536) & Total NFkB Cellular Assay KitsPhospho-p38 MAPK (Thr180/Tyr182) Cellular Assay KitPhospho-P70S6K (Thr389) Cellular Assay Kit Phospho-S6RP (Ser235/236) Cellular Assay Kit Phospho-STAT1 (Tyr701) & Total STAT1 Cellular Assay KitPhospho-STAT3 (Tyr705) & Total STAT3 Cellular Assay KitsPhospho-STAT5 (Tyr694) Cellular Assay KitPhospho-YAP (Ser127) & Total YAP Cellular Assay KitsPhospho-ERK (Thr202/Tyr204), Advanced phospho-ERK (Thr202/Tyr204), and Total ERK1/2 Cellular Assay KitsPhospho-SMAD1 (Ser463/465) & Total SMAD1 Cellular Assay KitsPhospho-p53 (Ser15) & Total p53 Cellular Assay KitsPhospho- & total AKT 1/2/3, Phospho- & total pan AKT Cellular Assay KitsPhospho-FGFR2 (Tyr653/654) & Total FGFR2 Cellular Assay KitsPhospho-MNK (Thr197/202) Cellular Assay KitPhospho-ULK1 (Ser757 mouse / Ser758 human) Assay KitTotal IRS2 Cellular Assay KitPhospho-SLP-76 (Ser376) & total SLP-76 Cellular Assay KitsPhospho-FoxO1 (Ser256) and Total FoxO1 Assay KitsPhospho-c-Src (Tyr419) and Total c-Src Assay KitsPhospho-IRF3 (Ser386) Cellular Assay KitPhospho-SMAD3 (Ser423/425) & Total SMAD3 Cellular Assay KitsPhospho-EIF2 alpha (Ser52) & Total EIF2 alpha Cellular Assay Kits