{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Woodland B"],"funding":["Natural Sciences and Engineering Research Council"],"pagination":["10"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC10059017"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["11(1)"],"pubmed_abstract":["Integrative top-down proteomics is an analytical approach that fully addresses the breadth and complexity needed for effective and routine assessment of proteomes. Nonetheless, any such assessments also require a rigorous review of methodology to ensure the deepest possible quantitative proteome analyses. Here, we establish an optimized general protocol for proteome extracts to improve the reduction of proteoforms and, thus, resolution in 2DE. Dithiothreitol (DTT), tributylphosphine (TBP), and 2-hydroxyethyldisulfide (HED), combined and alone, were tested in one-dimensional SDS-PAGE (1DE), prior to implementation into a full 2DE protocol. Prior to sample rehydration, reduction with 100 mM DTT + 5 mM TBP yielded increased spot counts, total signal, and spot circularity (i.e., decreased stre"],"journal":["Proteomes"],"pubmed_title":["Optimized Proteome Reduction for Integrative Top-Down Proteomics."],"pmcid":["PMC10059017"],"funding_grant_id":["2019-04324"],"pubmed_authors":["Coorssen JR","Woodland B","Necakov A"],"additional_accession":[]},"is_claimable":false,"name":"Optimized Proteome Reduction for Integrative Top-Down Proteomics.","description":"Integrative top-down proteomics is an analytical approach that fully addresses the breadth and complexity needed for effective and routine assessment of proteomes. Nonetheless, any such assessments also require a rigorous review of methodology to ensure the deepest possible quantitative proteome analyses. Here, we establish an optimized general protocol for proteome extracts to improve the reduction of proteoforms and, thus, resolution in 2DE. Dithiothreitol (DTT), tributylphosphine (TBP), and 2-hydroxyethyldisulfide (HED), combined and alone, were tested in one-dimensional SDS-PAGE (1DE), prior to implementation into a full 2DE protocol. Prior to sample rehydration, reduction with 100 mM DTT + 5 mM TBP yielded increased spot counts, total signal, and spot circularity (i.e., decreased stre","dates":{"release":"2023-01-01T00:00:00Z","publication":"2023 Mar","modification":"2025-04-22T18:19:35.559Z","creation":"2025-02-19T01:13:08.03Z"},"accession":"S-EPMC10059017","cross_references":{"pubmed":["36976889"],"doi":["10.3390/proteomes11010010"]}}