Simplified pathway dissection with HTRF phospho-assays and CyBi-FeliX liquid handling

2015

Cisbio Bioassays, Codolet, France

Higher throughput paired with reproducible and accurate results have become a “must” in these tough times of budget cuts and the ever-greater need to fight attrition. To meet this challenge, Cisbio cell signaling assay solutions and our partner CyBio instruments from Analytik Jena have combined their expertise to develop a flexible system to run Cisbio’s reliable, sensitive and rapid HTRF® phospho-protein assays on the CyBi®- FeliX liquid handling system for simplified pathway dissection. Here, the dissection of the PI3K/AKT/mTor translational control pathway is shown on different vertical signaling levels. Inhibitory effects of several compounds are measured, enabling their precise classification by elaborating a pathway map of their phospho-signatures. This application note demonstrates that Cisbio HTRF® assays and CyBio liquid handling form a perfect match offering a straightforward and flexible automated assay platform for drug discovery and cell signaling research.

Phosphorylated proteins

Phospho-P70S6K (Thr389) Cellular Assay Kit Phospho-4EBP1 (Thr37/46) Cellular Assay KitPhospho-EIF4E (Ser209) Cellular Assay Kit Phospho-S6RP (Ser235/236) Cellular Assay Kit Phospho-mTOR (Ser2448) 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-ERK (Thr202/Tyr204), Advanced phospho-ERK (Thr202/Tyr204), and Total ERK1/2 Cellular Assay KitsPhospho-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-NFkB (Ser536) & Total NFkB Cellular Assay KitsPhospho-p38 MAPK (Thr180/Tyr182) Cellular Assay KitPhospho-STAT3 (Tyr705) & Total STAT3 Cellular Assay KitsPhospho-STAT5 (Tyr694) Cellular Assay KitPhospho-YAP (Ser127) & Total YAP 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 KitsAlpha-Tubulin Housekeeping Cellular KitSTAT1 phospho (Tyr701) & total assaysPhospho-TBK1 (Ser172) & Total TBK1 Assay KitsPhospho-PDGFR beta (Tyr751) Cellular Assay Kit