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Immunoprecipitation of IL-6 from human serum for next-generation protein sequencing on Platinum

Quantum-Si
23 Jan 2024

In this application note, the focus is on enriching low-abundant proteins from complex biological matrices in proteomic workflows. The technical note introduces an immunoprecipitation protocol compatible with Quantum-Si's next-generation protein sequencing workflow. This protocol involves an on-bead digestion approach, eliminating the sample elution step and proceeding directly to the preparation of digested peptide libraries. The feasibility of this method is demonstrated through the specific enrichment of IL-6 via immunoprecipitation from human serum, followed by protein sequencing and identification on Quantum-Si's Platinum® Next-Generation Protein Sequencer™.

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Platinum® Next-Generation Protein Sequencer™

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Easy-to-use, accessible benchtop single-molecule protein sequencing platform that empowers any lab, anywhere to obtain critical proteomic insights quickly and efficiently from the convenience of your lab, no expertise or infrastructure required.

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ProteomicsProteomics is the systemic bioinformatics study of proteins and amino acids, including their structure, size, function and identification. Tools used in proteomics include chromatography, blotting and gels, protein arrays, mass spectrometry and ELISA and associated analysis software. Analyzers and proteomic systems should be sensitive, high resolution, fast and may be automated for high-throughput.BiomarkersBiomarkers are biological markers which can be measured and evaluated to indicate a biological state. The use of biomarkers in research and diagnosis can indicate a normal or disease state or drug response of cells / tissues. Biomarkers include genetic markers, cell surface markers such as antigens, antibodies or receptors and secreted molecules such as cytokines. An assay system is required for identification of biomarkers. :Next Generation SequencingNext-generation sequencing (NGS), also known as whole-genome sequencing, high-throughput sequencing and massive parallel sequencing, produces and analyses thousands to millions of nucleotide sequences at once. Sequencing systems operate via varying technologies depending on the manufacturer, including sequencing by synthesis, ligation, pyrosequencing, ion semiconductor and single-molecule real-time sequencing. For NGS, library preparation is paramount to successful sequencing. In this section, explore a range of library preparation kits, from targeted, amplicon-based or hybridization-based kits including epigenomic, transcriptomic and genomic workflows to fragmentation kits. Find the best next-generation sequencing products in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.ImmunoprecipitationSequencingSequencing refers to determining the order of nucleotides in a DNA or RNA molecule. This process is essential in genomics, diagnostics, and evolutionary biology. Techniques like next-generation sequencing (NGS) and Sanger sequencing provide high-throughput and precise data for analyzing genomes and transcriptomes. Browse our peer-reviewed product directory to find the best sequencing tools and reagents, compare products, check reviews, and get pricing directly from manufacturers.Protein BiologyThe analysis of protein expression, identity and function is vital for many areas of life science research and drug discovery. Some of the most commonly used techniques in protein analysis include Western blotting, electrophoresis and mass spectrometry.

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Immunoprecipitation of IL-6 from human serum for next-generation protein sequencing on Platinum