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Thermo Scientific™ SureQuant™ IP-MS Sample Preparation Kits
Description
Thermo Scientific SureQuant IP-MS Sample Preparation kits (Protein A/G or streptavidin) provide an optimized solution for the immunoprecipitation and MS peptide preparation of samples upstream of LC-MS/MS analysis. These kits have been rigorously verified using numerous multiplexed target antigens with varying expression levels. The SureQuant product family enables the use of the immunoprecipitation (IP) sample prep kits and peptide quantitation modules together or independently for SRM/PRM analysis.
• Multiplex—IP-MS kits have been designed for multiplex IP using both Protein A/G and Streptavidin magnetic beads
• Flexible—universal reagents enable individualized selection of targets for IP to be used alone or combined with SureQuant peptide quantitation panels
• Optimized—simplified and condensed procedure to prepare samples for LC-MS analysis
• Low background—binding, wash, and elution buffers optimized to remove contaminants and minimize enrichment of background proteins
The SureQuant Protein A/G and Streptavidin IP-MS Sample Preparation kits have been designed to support targeted quantitation using SureQuant or SRM/PRM analysis. The kits contain high-quality Pierce Protein A/G or Streptavidin magnetic beads together with reagents that have been optimized for MS compatibility. The streamlined procedure enables digestion immediately after the immunoprecipitation elution step, facilitating MS sample preparation in ∼4 hours for same-day LC-MS/MS analysis.
Applications:
• Enrichment of low abundance targets to improve MS sensitivity
• Identification of target proteins that are not detectable by western blotting
• Quantitation of target proteins by LC-MS analysis
• Screening of antibodies for IP-MS
• Enrichment of target proteins upstream of targeted analysis (SRM)
Specifications
Specifications
| Description | Streptavidin Magnetic Beads |
| Kit Contents | Trypsin Protease, 2 x 20 μg Trypsin Storage Solution, 250 μL Streptavidin Magnetic Beads, 1 mL IP-MS Cell Lysis Buffer, 100 mL IP-MS Wash Buffer A, 75 mL IP-MS Wash Buffer B, 40 mL IP Elution and MS Sample Prep Buffer, 10 mL Alkylation Solution, 1 mL/vial Low Protein-Binding Collection Tubes, 50 x 1.5 mL Bond-Breaker™ TCEP Solution, Neutral pH, 0.5 mL 10% Pierce™ Trifluoroacetic Acid (TFA), Sequencing Grade, 0.5 mL |
| For Use With (Equipment) | Mass spectrometer |
| Product Line | SureQuant |
| Content And Storage | Store trypsin at 4°C. Store trypsin storage solution at -20°C. |
| Starting Material | Cells, Tissue |
| Detection Method | Mass Spectrometry |
| Workflow Step | Sample prep enrichment |
| Quantity | 20 Reactions |
| For Use With (Application) | MS-compatible immunoprecipitation |
Frequently Asked Questions (FAQs)
SureQuant Protein IP-MS Sample Preparation kits use a trypsin enzyme elution method to dissociate the bound immune complexes from the magnetic beads, which can then be digested directly in-solution and followed up by MS analysis on the same day; no SDS-PAGE purification is required. The entire process take 4 hours compared to traditional IP methods that usually take around 21 hrs.
SureQuant IP-MS Sample Preparation kits include all reagents necessary for sample lysis, immunoenrichment, elution, digestion, and MS analysis. The entire process is completed in 4 hr for same-day LC-MS/MS analysis as compared to traditional IP methods that usually take around 21 hr. Pierce MS-Compatible Magnetic IP kits provide reagents necessary for the lysis step and the elution step, but not for the digestion step prior to MS analysis.
SureQuant IP-MS sample preparation kits have been designed for use with SureQuant quantitation modules for upstream sample preparation for SRM/PRM analysis, but they can be used with any samples where the target protein needs to be enriched by IP or IP-CoIP, prior to MS analysis.