Optimize your HTRF cell signaling assays for tissues

2018

Cisbio, Codolet, France

HTRF Cell Signaling assays allow rapid and reliable measurement of the phosphorylation and the expression level of key signaling proteins in cell lysates. These assays are also suitable for tissue sample analysis to evaluate the in vivo efficacy of compounds on preclinical models. However, the proper preparation of lysates and working solutions using the right reagents is crucial to ensure you obtain the best results. Moreover, it is essential to include the appropriate controls to analyze and interpret the results correctly. This technical note provides procedures and guidelines to optimize your HTRF Cell Signaling assays on tissues, with a focus on sample preparation and data analysis. Please note that the procedure for tissue lysis and sample preparation is also suitable for western-blot experiments.

Kinases, Phosphorylated proteins

Phospho-4EBP1 (Thr37/46) Cellular Assay KitPhospho-ACC (Ser79) Cellular Assay KitPhospho- & total AKT 1/2/3, Phospho- & total pan AKT Cellular Assay KitsPhospho-AMPK (Thr172) & Total AMPK Cellular Assay Kits Phospho-ATF2 (Thr71) Cellular Assay KitPhospho-Bad (Ser112) Cellular Assay Kit Phospho-BTK (Tyr223) & Total BTK Cellular Assay KitsPhospho-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-Insulin Receptor (Tyr1150/1151) & Total Insulin Receptor Cellular Assay KitsPhospho-IRS1 (Ser307 mouse / Ser312 human) & Total IRS1 Cellular Assay KitsPhospho-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-p53 (Ser15) & Total p53 Cellular Assay KitsPhospho-P70S6K (Thr389) Cellular Assay Kit Phospho-PDGFRβ (Tyr751) Cellular Assay KitPhospho-Phospholamban (Thr17) & total Phospholamban Cellular Assay KitsPhospho-S6RP (Ser235/236) Cellular Assay Kit Phospho-SMAD1 (Ser463/465) & Total SMAD1 Cellular Assay KitsPhospho-STAT3 (Tyr705) & Total STAT3 Cellular Assay KitsPhospho-STAT5 (Tyr694) Cellular Assay KitPhospho-TAU (Ser202/Thr205) Cellular Assay KitPhospho-VEGFR2 (Tyr1175) Cellular Assay KitPhospho-YAP (Ser127) & Total YAP Cellular Assay KitsPhospho-β-Catenin (Thr41/Ser37/Ser33) & Total β-Catenin Cellular Assay Kits Phospho-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-ZAP-70 (Tyr319) & Total ZAP-70 Cellular Assay KitsPhospho-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-VASP (Ser157), Phospho-VASP (Ser239) & Total VASP Cellular Assay KitsPhospho-SMAD3 (Ser423/425) & Total SMAD3 Cellular Assay KitsPhospho-EIF2 alpha (Ser52) & Total EIF2 alpha Cellular Assay KitsAlpha-Tubulin Housekeeping Cellular KitSTAT1 phospho (Tyr701) & total assays