{"doi":"10.3791/63602-v","title":"Purification of Ubiquitinated p53 Proteins from Mammalian Cells","abstract":null,"journal":"Journal of Visualized Experiments","year":2022,"id":676980,"datarank":0.10397207708399181,"base_score":0.6931471805599453,"endowment":0.6931471805599453,"self_citation_contribution":0.10397207708399181,"citation_network_contribution":0.0,"self_endowment_contribution":0.10397207708399181,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":1,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1768795,"name":"Mengfan He","orcid":null,"position":1,"is_corresponding":false},{"id":669062,"name":"Dawei Li","orcid":"0000-0002-3266-5272","position":2,"is_corresponding":false},{"id":495234,"name":"Di Wu","orcid":"0000-0002-3238-6388","position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"Purification of Ubiquitinated p53 Proteins from Mammalian Cells","abstract":"Ubiquitination is a type of posttranslational modification that regulates not only the stability but also the localization and function of a substrate protein. The ubiquitination process occurs intracellularly in eukaryotes and regulates almost all basic cellular biological processes. Purification of ubiquitinated proteins aids the investigation of the role of ubiquitination in controlling the function of substrate proteins. Here, a step-by-step procedure to purify ubiquitinated proteins in mammalian cells is described with the p53 tumor suppressor protein as an example. Ubiquitinated p53 proteins were purified under stringent nondenaturing and denaturing conditions. Total cellular Flag-tagged p53 protein was purified with anti-Flag antibody-conjugated agarose under nondenaturing conditions. Alternatively, total cellular His-tagged ubiquitinated protein was purified using nickel-charged resin under denaturing conditions. Ubiquitinated p53 proteins in the eluates were successfully detected with specific antibodies. Using this procedure, the ubiquitinated forms of a given protein can be efficiently purified from mammalian cells, facilitating studies on the roles of ubiquitination in regulating protein function.","is_dataset_classified":null,"base_score":0.6931471805599453,"endowment":0.6931471805599453,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"26207759","pmcid":null,"openalex_id":"https://openalex.org/W4327806861","authors":[],"funders":[],"total_grants":0,"fwci":0.0931,"citation_percentile":0.45925438,"influential_citations":0,"citation_trend":[{"year":2023,"count":1}],"oa_status":"closed","license":null,"oa_locations":[{"url":"https://www.jove.com/v/63602/purification-of-ubiquitinated-p53-proteins-from-mammalian-cells","host_type":"publisher"},{"url":"https://doi.org/10.3791/63602-v","host_type":"journal"}],"fields_of_study":["Cancer-related Molecular Pathways","Ubiquitin and proteasome pathways","Cancer Research and Treatments"],"mesh_terms":[],"keywords":["Ubiquitin","F-box protein","Cell biology","Agarose","Biology","Ubiquitin-Protein Ligases","Ubiquitins","Function (biology)","Biochemistry","Molecular biology","Chemistry","Ubiquitin ligase","Gene"],"sdg_mappings":[],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[],"source":"live","citation_network_status":"fetched"},"created_at":"2026-08-17T03:26:07.230779Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}