AKT PHOSPHO MULTIPLEX ABSOLUTE 10 RXNS，A40084，赛默飞世尔

The Thermo Scientific SureQuant AKT Pathway (Phospho) Multiplex Panel (Absolute Quantitation) provides a complete reagent and quantitation solution for the enrichment and analysis of key phosphorylation events of AKT signaling pathway proteins using mass spectrometry (MS).

The SureQuant AKT Pathway (Phospho) Multiplex Panel (Absolute Quantitation) consists of two modules:
• SureQuant AKT Pathway (Phospho) IP and MS Sample Preparation Module includes reagents necessary to immuno-enrich phosphorylated AKT-mTOR pathway proteins and perform in-solution MS sample preparation from sample lysis through peptide clean-up
• SureQuant AKT Pathway (Phospho) Absolute Quantitation Module includes a system suitability standard to monitor MS performance, AQUA Ultimate HeavyPeptide and LightPeptide mixtures for absolute peptide quantitation, and methods for using Thermo Scientific Q Exactive and Fusion Tribrid instruments and Nanospray LC instrumentation

SureQuant AKT Pathway (Phospho) Multiplex Panel features include:
• Complete—includes all reagents for successful monitoring of MS system performance, sample preparation, and absolute quantitation of target proteins/peptides
• Validated—antibodies, peptides, and control lysate rigorously tested for specificity and successful quantitation of each target peptide
• Multiplex—able to quantitate phosphorylation of ten key proteins in the AKT signaling pathway
• Flexible—modular format allows for immuno-enrichment only, or in combination with relative or absolute quantitation

The AKT signaling pathway serves as a master regulatory pathway for cell growth and proliferation, and phosphorylation plays a major role in its modulation. Aberrant activation of this pathway promotes survival of tumor cells in many types of cancer. This multiplex kit is ideal for monitoring protein expression of ten key phosphorylated proteins in the AKT signaling pathway (AKT, PTEN, IRS1, IGF1-R, GSK3a, GSK3b, PRAS40, mTOR, p70S6K, and TSC2) in a single assay. The ability to simultaneously monitor and quantitate these targets in a single sample is a powerful tool for assessment of normal and abnormal pathway signaling.

The SureQuant AKT Pathway (Phospho) IP and MS Sample Preparation Module enables highly effective antigen immunoprecipitation (IP) and co-immunoprecipitation (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, with non-specific proteins removed through washing with MS-compatible buffers. The target proteins are eluted using trypsin, and are directly compatible with MS sample preparation. The MS sample preparation has been streamlined and can be completed in ~4 hours for same-day MS analysis.

The SureQuant AKT Pathway (Phospho) Absolute Quantitation Module provides reagents for absolute quantitation of 30 unique peptides from 10 phosphorylated AKT-mTOR signaling pathway target proteins using liquid chromatography (LC) and MS. AQUA Ultimate HeavyPeptide mix, AQUA Ultimate LightPeptide mix, and Pierce 6 Protein Digest, equimolar, LC-MS grade reagents are used to generate a standard curve in a matrix, and serial dilutions performed by the user have been optimized to span 2–3 orders of magnitude (200 to 0.03 fmol on column). Standardized methods and data process templates are provided for streamlined data acquisition and analysis.

The Pierce LC-MS/MS System Suitability Standard (7 x 5 Mix) is included in this module to assess dynamic range of LC-MS/MS systems before running the calibration curves and samples.

Together, these modules provide a complete solution for sample preparation and analysis of the phosphorylated target proteins in the pathway.

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SureQuant AKT Pathway Multiplex Panel (Absolute Quantitation)
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