HTRF phospho- total lysis buffer: a universal alternative to RIPA lysis buffers

2015

Cisbio Bioassays, Codolet, France

Cisbio offers a comprehensive line of HTRF phospho- and total protein assays to investigate cell signaling pathways. These versatile, homogeneous assays are suitable for a variety of sample types, including primary cell lines and tumor xenografts. Each HTRF assay includes a common and essential step that relies on the ability of the lysis buffer to release the protein of interest. This protein, either phosphorylated or not, is then detected by an HTRF sandwich immunoassay.
This application note compares the use of Cisbio’s HTRF phospho-total lysis buffer with commonly used RIPA lysis buffers for the analysis of cell signaling pathways in different types of samples. The data demonstrate that HTRF phospho-total lysis buffer is as efficient as RIPA lysis buffers to perform protein extraction and is compatible with both simple and more complex biological samples. Moreover, the HTRF phospho-total lysis buffer delivers higher quality HTRF results than RIPA lysis buffers. Finally the HTRF phospho-total Cisbio lysis buffer is not only suitable for HTRF, but also for other analytical methods such as Western blot and ELISA.

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-TAU (Ser202/Thr205) 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-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