{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T11:41:49Z","timestamp":1788349309867,"version":"build-2803163510"},"reference-count":30,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2018,10,30]],"date-time":"2018-10-30T00:00:00Z","timestamp":1540857600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100003196","name":"Ministero della Salute","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100003196","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["currentprotocols.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["CP Molecular Biology"],"published-print":{"date-parts":[[2019,1]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>In the past few decades, numerous approaches have been developed to investigate protein\u2010protein and protein\u2010nucleic acid interactions (PPIs and PNIs). Affinity purification methods such as co\u2010immunoprecipitation (co\u2010IP) are commonly used to detect and isolate the macromolecular complexesresulting from these interactions. In this article, we describe a two\u2010step co\u2010immunoprecipitation (TIP) technique. As compared to standard co\u2010IP, TIP provides increased specificity in the isolation of PPIs or PNIs under native expression conditions, dramatically reducing the abundance of nonspecific binders and thus facilitating downstream analyses of the interaction complexes. Here, we report a detailed TIP procedure that we used to purify a protein\u2010protein complex from Burkitt lymphoma cells and from primary human CD4<jats:sup>+<\/jats:sup> T cells. In addition, this unit describes an application of TIP for the isolation of transcription\u2010factor\u2010bound chromatin. \u00a9 2018 by John Wiley &amp; Sons, Inc.<\/jats:p>","DOI":"10.1002\/cpmb.80","type":"journal-article","created":{"date-parts":[[2018,10,30]],"date-time":"2018-10-30T14:56:35Z","timestamp":1540911395000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Two\u2010Step Co\u2010Immunoprecipitation (TIP)"],"prefix":"10.1002","volume":"125","author":[{"given":"Maria Rita","family":"Sciuto","sequence":"first","affiliation":[{"name":"Department of Oncology and Molecular Medicine, Istituto Superiore di Sanit\u00e0  Rome Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Valeria","family":"Coppola","sequence":"additional","affiliation":[{"name":"Department of Oncology and Molecular Medicine, Istituto Superiore di Sanit\u00e0  Rome Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gioacchin","family":"Iannolo","sequence":"additional","affiliation":[{"name":"Regenerative Medicine and Biomedical Technologies Unit, Department of Laboratory Medicine and Advanced Biotechnologies, IRCCS\u2010ISMETT  Palermo Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruggero","family":"De Maria","sequence":"additional","affiliation":[{"name":"Institute of General Pathology, Universit\u00e0 Cattolica del Sacro Cuore  Rome Italy"},{"name":"Fondazione Policlinico Universitario A. 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Gemelli IRCCS  Rome Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2018,10,30]]},"reference":[{"key":"e_1_2_8_2_1_1","doi-asserted-by":"publisher","DOI":"10.1038\/521274a"},{"key":"e_1_2_8_2_2_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1105776"},{"key":"e_1_2_8_2_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymeth.2015.08.022"},{"issue":"4","key":"e_1_2_8_2_4_1","doi-asserted-by":"crossref","first-page":"1283","DOI":"10.4049\/jimmunol.143.4.1283","article-title":"Mechanisms of T cell activation by the calcium ionophore ionomycin","volume":"143","author":"Chatila T.","year":"1989","journal-title":"Journal of Immunology (Baltimore, Md.: 1950)"},{"key":"e_1_2_8_2_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/0378-1119(93)90262-2"},{"key":"e_1_2_8_2_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0165-022X(01)00213-5"},{"key":"e_1_2_8_2_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/0008-8749(84)90141-2"},{"key":"e_1_2_8_2_8_1","doi-asserted-by":"publisher","DOI":"10.1002\/0471142727.mb1008s114"},{"key":"e_1_2_8_2_9_1","doi-asserted-by":"publisher","DOI":"10.1038\/415141a"},{"key":"e_1_2_8_2_10_1","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.2534"},{"key":"e_1_2_8_2_11_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrm2208"},{"key":"e_1_2_8_2_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2008.01.020"},{"key":"e_1_2_8_2_13_1","doi-asserted-by":"publisher","DOI":"10.1002\/0471142735.im0503s44"},{"key":"e_1_2_8_2_14_1","doi-asserted-by":"publisher","DOI":"10.1101\/cshperspect.a000158"},{"key":"e_1_2_8_2_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2016.10.044"},{"key":"e_1_2_8_2_16_1","doi-asserted-by":"publisher","DOI":"10.1042\/BA20090273"},{"key":"e_1_2_8_2_17_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF00986726"},{"key":"e_1_2_8_2_18_1","doi-asserted-by":"crossref","first-page":"3.14.11","DOI":"10.1002\/cptx.22","article-title":"Using chromatin immunoprecipitation in toxicology: A step\u2010by\u2010step guide to increasing efficiency, reducing variability, and expanding applications","volume":"72","author":"McCullough S. 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R. Warnken U. Schnolzer M. Valvo C. Brunetto L. Boe A. \u2026Haas T. L.(2018). See above."}],"container-title":["Current Protocols in Molecular Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fcpmb.80","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cpmb.80","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,9,14]],"date-time":"2023-09-14T09:16:36Z","timestamp":1694682996000},"score":1,"resource":{"primary":{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/10.1002\/cpmb.80"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,10,30]]},"references-count":30,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2019,1]]}},"alternative-id":["10.1002\/cpmb.80"],"URL":"https:\/\/doi.org\/10.1002\/cpmb.80","archive":["Portico"],"relation":{},"ISSN":["1934-3639","1934-3647"],"issn-type":[{"value":"1934-3639","type":"print"},{"value":"1934-3647","type":"electronic"}],"subject":[],"published":{"date-parts":[[2018,10,30]]},"assertion":[{"value":"2018-10-30","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e80"}}