Description
More Peptides. More Coverage. More Certainty.
With insights beyond proline, expanded amino acid recognition and added precision:
- Characterize proteins
- Sequence antibodies
- Resolve mixtures
Unlock Proteome Depth with the Sequencing Kit V4
Single-molecule resolution to identify and characterize proteins on a benchtop instrument
Insights Beyond Proline
Navigate challenges posed by proline-rich domains, common in antibodies and membrane proteins, with Sequencing Kit V4
Expanded Coverage
New recognizers, peptidases, and improved bioinformatics offer an 85% increase in sequenceable peptides
Protein Mixture Precision
Enhanced bioinformatics to identify peptides with greater confidence.
Sequencing Through Proline
- V4 detects amino acids beyond proline, which accounts for 6% of the total amino acids in the human proteome and over 40% of peptides generated by Lys-C digestion
- Proline is abundant in vital proteins that are difficult to analyze by mass spec, common in membrane proteins, antibodies, and transcription factors
Proline’s unique structure impacts the peptide fragmentation for mass spec analysis, a phenomenon referred to as the “proline effect,” which restricts sequencing by mass spec. The V4 kit delivers improved chemistry that sequences through proline rich regions, expanding sequencing insights on Platinum.
Expanded Proteome Coverage
A panel of ten recombinant proteins were digested into peptides, and sequenced using the Platinum Pro with both the V4 and V3 kits demonstrating:
- Increased peptides coverage in 8 of 10 proteins
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Increased peptide alignments (sequenced copies) in 10 of 10 proteins
Protein Mixture Precision
Upgrades to the analysis software boost confidence in accurate protein identification in mixtures using V4. An equimolar mixture of ten proteins sequenced with V4 demonstrated increased peptide alignments, resulting in increased precision in peptides identified compared to V3.
Sequence Antibodies
Prolines play a crucial role in antibody structure and function, shaping complementarity-determining regions (CDRs) and immunoglobulin loops. We sequenced trastuzumab, an anti-HER2 monoclonal antibody used in cancer treatment, on the Platinum Pro, to demonstrate the improved coverage obtainable with V4, exhibiting:
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2.5-fold more peptide coverage in heavy and light chains compared to V3
- Significant increase in amino acid calls after proline with V4 chemistry
Characterize Proteins by Immunoprecipitation
Traditional methods for identification of proteins enriched by immunoprecipitation (IP) vary in their resolution, where Western blot offers qualitative to semi-quantitative measurements that are antibody-dependent, and mass spectrometry can produce exhaustive data inclusive of proteins that may be nonessential to a study. Platinum Pro with the V4 kit is an accessible non-antibody-dependent, higher-resolution option optimized to identify target proteins with concise data readouts.
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Sequence IP products on Platinum Pro for clear identification of protein interactors
- Explore signaling pathways, antibody binding partners, and other protein interactions
