HTRF Phospho-assays reveal subtle drug-induced effects in tumor xenografts. A method of choice beyond western blot

2015

Cisbio Bioassays, Codolet, France

Cisbio offers a comprehensive line of HTRF phospho- and total protein assays to study posttranslational cellular modifications. These versatile, homogeneous assays are suitable for a variety of sample types, including primary cell lines and tumor xenografts. Combining an HTRF phospho-assay with its associated total protein assay enables researchers to accurately identify candidate molecules that regulate phosphorylation, as well as to characterize their mechanism of action by performing the appropriate data analysis.
This application note compares the use of HTRF phospho- and total protein assays with traditional Western blot for analyzing cell signaling pathways in tumor xenografts. The data demonstrates that HTRF is a faster, simpler, and more accurate technique for assessing protein phosphorylation and for revealing subtle drug-induced modulations than Western blotting.

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-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-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-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