{"doi":"10.1002/0471142735.im1104s07","title":"Immune‐Complex Assays for Tyrosine Protein Kinases","abstract":"AbstractTyrosine protein kinases (TPKs) represent a diverse group of enzymes that contribute to cellular signal transduction. The generally low abundance of TPKs, coupled with their rapid activation and deactivation, usually precludes their purification through conventional biochemical means. Using immune‐complex protein kinase assays, the presence or absence of a given TPK can be established and an estimation of its functional state obtained. In the of this unit, TPKs are immunoprecipitated, allowed to autophosphorylate in the presence of labeled ATP, run out on an SDS‐PAGE gel, and detected by autoradiography. Alternate protocols are provided for the assessment of the functional state of TPKs by providing a potential substrate along with the labeled ATP in the reaction mixture. In the first alternate protocol, the exogenous substrate is a protein, permitting simultaneous assessment of autophosphorylation and exogenous substrate phosphorylation. The second alternate protocol utilizes a peptide substrate, resulting in a rapid, high‐throughput assay that evaluates only exogenous substrate phosphorylation.","journal":"Current Protocols in Immunology","year":1993,"id":673833,"datarank":0.40620753016533157,"base_score":2.70805020110221,"endowment":2.70805020110221,"self_citation_contribution":0.40620753016533157,"citation_network_contribution":0.0,"self_endowment_contribution":0.40620753016533157,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":14,"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":1760563,"name":"Joseph B. Bolen","orcid":null,"position":1,"is_corresponding":false},{"id":1760561,"name":"Anne L. Burkhardt","orcid":null,"position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"Immune‐Complex Assays for Tyrosine Protein Kinases","abstract":"AbstractTyrosine protein kinases (TPKs) represent a diverse group of enzymes that contribute to cellular signal transduction. The generally low abundance of TPKs, coupled with their rapid activation and deactivation, usually precludes their purification through conventional biochemical means. Using immune‐complex protein kinase assays, the presence or absence of a given TPK can be established and an estimation of its functional state obtained. In the of this unit, TPKs are immunoprecipitated, allowed to autophosphorylate in the presence of labeled ATP, run out on an SDS‐PAGE gel, and detected by autoradiography. Alternate protocols are provided for the assessment of the functional state of TPKs by providing a potential substrate along with the labeled ATP in the reaction mixture. In the first alternate protocol, the exogenous substrate is a protein, permitting simultaneous assessment of autophosphorylation and exogenous substrate phosphorylation. The second alternate protocol utilizes a peptide substrate, resulting in a rapid, high‐throughput assay that evaluates only exogenous substrate phosphorylation.","is_dataset_classified":null,"base_score":2.70805020110221,"endowment":2.70805020110221,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"18432705","pmcid":null,"openalex_id":"https://openalex.org/W1556202419","authors":[],"funders":[],"total_grants":0,"fwci":0.8551,"citation_percentile":0.71945616,"influential_citations":0,"citation_trend":[{"year":2016,"count":1},{"year":2022,"count":1}],"oa_status":"closed","license":"http://onlinelibrary.wiley.com/termsAndConditions#vor","oa_locations":[{"url":"https://onlinelibrary.wiley.com/doi/pdf/10.1002/0471142735.im1104s07","host_type":"publisher"},{"url":"https://onlinelibrary.wiley.com/doi/full-xml/10.1002/0471142735.im1104s07","host_type":"publisher"},{"url":"https://currentprotocols.onlinelibrary.wiley.com/doi/pdf/10.1002/0471142735.im1104s07","host_type":"publisher"},{"url":"https://doi.org/10.1002/0471142735.im1104s07","host_type":"journal"},{"url":"https://pubmed.ncbi.nlm.nih.gov/18432705","host_type":"repository"}],"fields_of_study":["Monoclonal and Polyclonal Antibodies Research","Chronic Lymphocytic Leukemia Research"],"mesh_terms":["Adenosine Triphosphate","Animals","Autoradiography","Caseins","Electrophoresis, Polyacrylamide Gel","Humans","Aprotinin","Leupeptins","Peptides","Phenylmethylsulfonyl Fluoride","Phosphopyruvate Hydratase","Phosphorylation","Protein-Tyrosine Kinases","Rabbits","Sodium Fluoride","Structure-Activity Relationship","Time Factors","Vanadates","Signal Transduction","Cell Line, Tumor","Protein Kinase Inhibitors","Immunoprecipitation"],"keywords":["Autophosphorylation","Phosphorylation","Kinase","Immunoprecipitation","Substrate-level phosphorylation","Biochemistry","Protein phosphorylation","Substrate (aquarium)","Biology","Signal transduction","Tyrosine phosphorylation","Enzyme","Tyrosine kinase","Cell biology","Protein kinase A","Chemistry"],"sdg_mappings":[],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[],"source":"live","citation_network_status":"fetched"},"created_at":"2026-08-16T14:38:06.514261Z","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":[]}