{"doi":"10.1016/j.xpro.2025.104086","title":"Protocol for in vivo BONCAT labeling of nascent proteins in Xenopus laevis tadpole brains with high temporal precision for quantitative analysis","abstract":"Proteostasis is critical for neuronal function and circuit plasticity. We present here an in vivo bio-orthogonal non-canonical amino acid tagging (BONCAT) protocol optimized to selectively label newly synthesized proteins in Xenopus laevis tadpole brains in live animals. We describe steps for the quantitative analysis of both total nascent proteins and specific nascent proteins of interest, using dot blotting and NeutrAvidin purification followed by western blotting. This approach allows for the quantitative analysis of the nascent proteome in intact neural circuits with high temporal precision. For complete details on the use and execution of this protocol, please refer to He et al. 1 • Procedure for high-temporal precision in vivo protein labeling in live tadpole brain • Steps for quantitative evaluation of total nascent protein levels using dot blot assay • Guide for experimental design and validation for in vivo tagging of proteins via BONCAT Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Proteostasis is critical for neuronal function and circuit plasticity. We present here an in vivo bio-orthogonal non-canonical amino acid tagging (BONCAT) protocol optimized to selectively label newly synthesized proteins in Xenopus laevis tadpole brains in live animals. We describe steps for the quantitative analysis of both total nascent proteins and specific nascent proteins of interest, using dot blotting and NeutrAvidin purification followed by western blotting. This approach allows for the quantitative analysis of the nascent proteome in intact neural circuits with high temporal precision.","journal":"STAR Protocols","year":2025,"id":546976,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"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":0.9559,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2025-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":992150,"name":"Reshmi Bera","orcid":null,"position":1,"is_corresponding":false},{"id":1439425,"name":"Benjamin Baah Konadu","orcid":null,"position":2,"is_corresponding":false},{"id":563561,"name":"Yuan Gao","orcid":"0000-0003-2224-5538","position":3,"is_corresponding":false},{"id":815268,"name":"Hai‐yan He","orcid":"0000-0002-6692-6012","position":4,"is_corresponding":false},{"id":992149,"name":"Arifa Ahsan","orcid":null,"position":0,"is_corresponding":true}],"reference_count":18,"raw_metadata":null,"created_at":"2026-07-19T02:53:36.567932Z","pmid":"40956668","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":[]}