SureQuant™ AKT Isoform Relative Quantitation Module - FAQs

The absolute quantitation module contains both AQUA Ultimate HeavyPeptides Mixture and AQUA Ultimate LightPeptides Mixture. For the absolute quantitation module, the standard curve is generated by serially diluting the light peptides against a constant backdrop of heavy peptides in order to obtain a ratio. The matrix for dilution is the Pierce 6 Protein Digest Standard Equimolar, LC/MS Grade that also serves as a constant. The relative quantitation module contains only AQUA Ultimate HeavyPeptides Mixture as a positive control.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

The absolute quantitation module contains AQUA Ultimate HeavyPeptide and AQUA Ultimate LightPeptide mixtures for absolute peptide quantitation. The standard curve is generated by serially diluting the light peptides against a constant backdrop of heavy peptides in order to obtain a ratio. The matrix for dilution is the Pierce 6 Protein Digest Standard Equimolar, LC/MS Grade which also serves as a constant.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

With the SureQuant kits, we recommend using the Orbitrap Fusion Tribrid MS, or the Orbitrap Q Exactive series of MS instruments.

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A typical IP-MS workflow takes up to 21 hours whereas a SureQuant workflow takes around 4 hours.

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The proteins for SureQuant analysis are purified from samples using a cocktail of highly validated biotinylated antibodies that are premixed with the sample and then purified with streptavidin magnetic beads.

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The antibodies used in the panel for each pathway were selected for reactivity with multiple mammalian species (including but not limited to human, mouse and rat).

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We've tested only human cells and tissues, although these kits can theoretically be used with any mammalian cells or tissues. We've tested fresh and fresh-frozen samples, both normal and tumor cells and tissues.

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SureQuant kits enable multiplex immunoprecipitation to mass spectrometry (mIP-MS) for the simultaneous enrichment and quantitation (absolute or relative) of multiple total and phosphorylated proteins (13 in total, including 11 phosphorylated states) in the AKT/mTOR signaling pathway.

For each pathway, we offer i) a pathway-specific IP-MS sample preparation module which includes all the reagents necessary to immunoenrich AKT/mTOR pathway proteins and perform in-solution MS sample preparation from sample lysis through peptide clean-up and ii) a quantitation module (relative or absolute), which includes a system suitability standard to monitor MS performance and AQUA Ultimate Heavy and /or Light Peptide mixtures for peptide quantitation. For each pathway, we also offer iii) a multiplex panel which is a combo kit comprising both IP-MS sample preparation and quantitation modules. Two versions of each multiplex panel are offered with either a relative or absolute quantitation module.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.