Thermo Scientific™ SureQuant™ AKT Pathway Relative Quantitation Module

Description

• The SureQuant Quantitation Modules contain 30 unique peptides from 10 AKT-mTOR signaling pathway target proteins for multiplexed quantitation of unique peptides using LC and MS
• SureQuant kits utilize multiplex immunoprecipitation coupled with mass spectrometry for simultaneous enrichment and quantitation of target proteins in the AKT/mTOR signaling pathway
• Each multiplex panel contains two modules: 1) the IP and MS Sample Preparation Module, which includes all the reagents necessary to immunoenrich target proteins with subsequent in-solution MS sample preparation, and 2) an Absolute or Relative Quantitation Module, including a system suitability standard, and HeavyPeptide™ and LightPeptide™ AQUA Ultimate mixtures
• For relative quantitation, a HeavyPeptide AQUA Ultimate mixture is provided for use as a spike-in internal standard in the multiplexed sample
• For absolute quantitation, HeavyPeptide and LightPeptide AQUA Ultimate mixtures are provided with the Pierce™ 6 Protein Digest are used to generate a standard curve in a matrix
• The Pierce™ LC-MS/MS System Suitability Standard (7 x 5 mix) is included in both modules to assess the dynamic range of LC-MS/MS systems before running the calibration curves and samples
• The IP and MS Sample Preparation Module enables highly effective antigen IP and co-IP for MS analysis
• Cells are first lysed and incubated with an IP-MS–verified biotinylated antibody mixture
• The antibody–antigen complex is then captured on MS-compatible streptavidin magnetic beads, and nonspecific proteins are removed through washing with MS-compatible buffers
• The target proteins are eluted using trypsin and are compatible with MS sample preparation
• The MS sample preparation has been streamlined to be completed in 4 hours so that the sample can be analyzed by MS on the same day
• The AKT/mTOR signaling pathway serves as a master regulatory pathway for cell growth and proliferation and plays a vital role in the signaling cascade
• Aberrant activation of this pathway promotes survival of tumor cells in many types of cancer
• SureQuant kits are ideal for monitoring AKT/mTOR expression levels, isoforms, and phosphorylation status in a single assay
• The ability to simultaneously monitor and quantitate in a single sample is a powerful tool for assessment of normal and abnormal cell growth
• SureQuant AKT Pathway Multiplex Panels are: • Complete —includes all reagents for successful monitoring of MS system performance, sample preparation, and quantitation of target proteins and peptides • Verified —antibodies, peptides, and control lysate are rigorously tested for specificity and successful quantitation of each target peptide • Multiplex —able to quantitate different targets from the AKT/mTOR pathway, RAS, and p53 isoforms • Flexible —modular format allows for immune-enrichment only, or in combination with relative or absolute quantitation