{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T16:46:04Z","timestamp":1791218764065,"version":"4.1.0"},"reference-count":47,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2015,7,29]],"date-time":"2015-07-29T00:00:00Z","timestamp":1438128000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["The Plant Journal"],"published-print":{"date-parts":[[2015,8]]},"abstract":"<jats:title>Summary<\/jats:title><jats:p>Manganese (Mn) constitutes an essential co\u2010factor in the oxygen\u2010evolving complex of photosystem <jats:styled-content style=\"fixed-case\">II<\/jats:styled-content> (<jats:styled-content style=\"fixed-case\">PSII<\/jats:styled-content>). Consequently, Mn deficiency reduces photosynthetic efficiency and leads to changes in <jats:styled-content style=\"fixed-case\">PSII<\/jats:styled-content> composition. In order to study these changes, multiplexed protein assays are advantageous. Here, we developed a multiplexed antibody\u2010based assay and analysed selected <jats:styled-content style=\"fixed-case\">PSII<\/jats:styled-content> subunits in barley (<jats:italic>Hordeum vulgare<\/jats:italic> L.). A selection of antibodies were labelled with specific lanthanides and immunoreacted with thylakoids exposed to Mn\u00a0deficiency after western blotting. Subsequently, western blot membranes were analysed by laser ablation inductively coupled plasma mass spectrometry (<jats:styled-content style=\"fixed-case\">LA<\/jats:styled-content>\u2010<jats:styled-content style=\"fixed-case\">ICP<\/jats:styled-content>\u2010<jats:styled-content style=\"fixed-case\">MS<\/jats:styled-content>), which allowed selective and relative quantitative analysis via the different lanthanides. The method was evaluated against established liquid chromatography electrospray ionization tandem mass spectrometry (<jats:styled-content style=\"fixed-case\">LC<\/jats:styled-content>\u2010<jats:styled-content style=\"fixed-case\">ESI<\/jats:styled-content>\u2010<jats:styled-content style=\"fixed-case\">MS<\/jats:styled-content>\/<jats:styled-content style=\"fixed-case\">MS<\/jats:styled-content>) methods, based on\u00a0data\u2010dependent acquisition (<jats:styled-content style=\"fixed-case\">DDA<\/jats:styled-content>) and selected reaction monitoring (<jats:styled-content style=\"fixed-case\">SRM<\/jats:styled-content>). Manganese deficiency resulted in a general decrease in <jats:styled-content style=\"fixed-case\">PSII<\/jats:styled-content> protein abundances, an effect that was shown to be reversible upon Mn re\u2010supplementation. Specifically, the extrinsic proteins PsbP and PsbQ showed Mn\u2010dependent changes in abundances. Similar trends in the response to Mn deficiency at the protein level were observed when comparing <jats:styled-content style=\"fixed-case\">DDA<\/jats:styled-content>,<jats:styled-content style=\"fixed-case\"> SRM<\/jats:styled-content> and <jats:styled-content style=\"fixed-case\">LA<\/jats:styled-content>\u2010<jats:styled-content style=\"fixed-case\">ICP<\/jats:styled-content>\u2010<jats:styled-content style=\"fixed-case\">MS<\/jats:styled-content> results. A biologically important exception to this trend was the loss of PsbO in the <jats:styled-content style=\"fixed-case\">SRM<\/jats:styled-content> analysis, which highlights the necessity of validating protein changes by more than one technique. The developed method enables a higher number of proteins to be multiplexed in comparison to existing immunoassays. Furthermore, multiplexed protein analysis by <jats:styled-content style=\"fixed-case\">LA<\/jats:styled-content>\u2010<jats:styled-content style=\"fixed-case\">ICP<\/jats:styled-content>\u2010<jats:styled-content style=\"fixed-case\">MS<\/jats:styled-content> provides an analytical platform with high throughput appropriate for screening large collections of plants.<\/jats:p>","DOI":"10.1111\/tpj.12906","type":"journal-article","created":{"date-parts":[[2015,6,19]],"date-time":"2015-06-19T14:52:51Z","timestamp":1434725571000},"page":"555-565","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":21,"title":["A laser ablation <scp>ICP<\/scp>\u2010<scp>MS<\/scp> based method for multiplexed immunoblot analysis: applications to manganese\u2010dependent protein dynamics of photosystem <scp>II<\/scp> in barley (<i><scp>H<\/scp>ordeum vulgare <\/i><scp>L<\/scp>.)"],"prefix":"10.1111","volume":"83","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5019-6422","authenticated-orcid":false,"given":"Thomas Christian","family":"de Bang","sequence":"first","affiliation":[{"name":"Department of Plant and Environmental Sciences Copenhagen Plant Science Centre Faculty of Science University of Copenhagen Thorvaldsensvej 40 DK\u20101871 Frederiksberg C Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J\u00f8rgen","family":"Petersen","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology University of Southern Denmark Campusvej 55, DK\u20105230, Odense M, Denmark Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pai Rosager","family":"Pedas","sequence":"additional","affiliation":[{"name":"Department of Plant and Environmental Sciences Copenhagen Plant Science Centre Faculty of Science University of Copenhagen Thorvaldsensvej 40 DK\u20101871 Frederiksberg C Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adelina","family":"Rogowska\u2010Wrzesinska","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology University of Southern Denmark Campusvej 55, DK\u20105230, Odense M, Denmark Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ole Noerregaard","family":"Jensen","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology University of Southern Denmark Campusvej 55, DK\u20105230, Odense M, Denmark Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jan Kofod","family":"Schjoerring","sequence":"additional","affiliation":[{"name":"Department of Plant and Environmental Sciences Copenhagen Plant Science Centre Faculty of Science University of Copenhagen Thorvaldsensvej 40 DK\u20101871 Frederiksberg C Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Poul Erik","family":"Jensen","sequence":"additional","affiliation":[{"name":"Department of Plant and Environmental Sciences Copenhagen Plant Science Centre Faculty of Science University of Copenhagen Thorvaldsensvej 40 DK\u20101871 Frederiksberg C Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jay J.","family":"Thelen","sequence":"additional","affiliation":[{"name":"Christopher S. 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