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[{"key": "dc.contributor.advisor", "value": "Kainulainen, Heikki", "language": "", "element": "contributor", "qualifier": "advisor", "schema": "dc"}, {"key": "dc.contributor.advisor", "value": "Lehti, Maarit", "language": "", "element": "contributor", "qualifier": "advisor", "schema": "dc"}, {"key": "dc.contributor.author", "value": "Paajala, Janne", "language": null, "element": "contributor", "qualifier": "author", "schema": "dc"}, {"key": "dc.date.accessioned", "value": "2017-06-15T09:47:25Z", "language": "", "element": "date", "qualifier": "accessioned", "schema": "dc"}, {"key": "dc.date.available", "value": "2017-06-15T09:47:25Z", "language": "", "element": "date", "qualifier": "available", "schema": "dc"}, {"key": "dc.date.issued", "value": "2017", "language": null, "element": "date", "qualifier": "issued", "schema": "dc"}, {"key": "dc.identifier.other", "value": "oai:jykdok.linneanet.fi:1703949", "language": null, "element": "identifier", "qualifier": "other", "schema": "dc"}, {"key": "dc.identifier.uri", "value": "https://jyx.jyu.fi/handle/123456789/54519", "language": "", "element": "identifier", "qualifier": "uri", "schema": "dc"}, {"key": "dc.description.abstract", "value": "Paajala J. 2017. Sirt1-, sirt3- ja PGC-1\u03b1-proteiinien immunopresipitaatio rotan gastrocnemius- ja soleuslihaksesta magneettien avulla. Liikuntatieteellinen tiedekunta, Jyv\u00e4skyl\u00e4n yliopisto, liikuntafysiologian pro gradu -tutkielma, 70 s. \r\n\r\nSirt1-, sirt3- ja PGC-1\u03b1-proteiinit s\u00e4\u00e4telev\u00e4t muun muassa solujen mitokondrioiden m\u00e4\u00e4r\u00e4\u00e4 ja toimintaa. Koska metabolisen oireyhtym\u00e4n sairaudet ovat yhteydess\u00e4 mitokondrioiden \r\nvajaatoimintaan, on n\u00e4iden proteiinin biologisen merkityksen tunteminen t\u00e4rke\u00e4\u00e4. Tutkimusk\u00e4ytt\u00f6\u00e4 varten on jalostettu syntyper\u00e4lt\u00e4\u00e4n heikon (low capacity runner rat, LCR) ja korkean (high capacity runner rat, HCR) juoksukyvyn omaavat rottakannat, joiden yksil\u00f6t eroavat toisistaan terveydentilaltaan. HCR/LCR -mallin avulla voidaan tutkia, kuinka aineenvaihduntaan osallistuvien entsyymien toiminta eroaa terveill\u00e4 ja sairailla yksil\u00f6ill\u00e4. \r\n\r\nT\u00e4ss\u00e4 tutkimuksessa selvitettiin magnetiikkaan perustuvan immunopresipitaation avulla sirt1-, sirt3-, PGC-1\u03b1-, GAPDH- ja tubulin-proteiinien eristyst\u00e4. HCR-rotan homogenoituun \r\nlihasn\u00e4ytteeseen lis\u00e4ttiin ensin kohdeproteiinit tunnistavat vasta-aineet. Sen j\u00e4lkeen lis\u00e4ttiin vasta-aineisiin tarttuvat magneetit. Vaihtoehtoisesti ensin kiinnitettiin magneetit ja vasta-aine, mink\u00e4 j\u00e4lkeen lis\u00e4ttiin homogenaatti. Seuraavana p\u00e4iv\u00e4n\u00e4 magneetit ker\u00e4ttiin liuoksesta ja ne puhdistettiin siten, ett\u00e4 magneettiin kiinnittynyt vasta-aine ja kohdeproteiini irtosivat magneetista. Kohdeproteiini analysoitiin Western blot -menetelm\u00e4ll\u00e4. \r\n\r\nTulosten mukaan sirt1-, sirt3-, PGC-1\u03b1-, tubulin ja GAPDH-proteiineja ei pystytty erist\u00e4m\u00e4\u00e4n tutkimuksessa k\u00e4ytettyjen menetelmien avulla. Immunopresipitaatiossa k\u00e4ytetty vasta-aine irtosi magneetista kohdeproteiinin mukana Western blot -analyysiin, jossa se h\u00e4iritsi kuvantamista. Toinen ongelma oli se, ett\u00e4 vasta-aine ei tarttunut kiinni kohdeproteiiniinsa tai magneetti ei tarttunut vasta-aineeseen. Sen seurauksena osa kohdeproteiineista peseytyi immunopresipitaation aikana pois. Western blot -menetelm\u00e4ll\u00e4 PGC-1\u03b1-proteiinia ei havaittu. \r\n\r\nMuokkaamalla menetelm\u00e4n olosuhteita, kuten puskureiden pH-arvoja ja vaihtamalla k\u00e4ytett\u00e4vi\u00e4 vasta-aineita, menetelm\u00e4 saataneen onnistumaan. Lis\u00e4ksi erilaisia vasta-aineen ja magneetin kiinnitt\u00e4mistapoja on syyt\u00e4 kokeilla, sill\u00e4 vasta-aineen ja magneetin v\u00e4liin laitettavalla sitojakomponentilla voitaneen est\u00e4\u00e4 vasta-aineen p\u00e4\u00e4seminen kohdeproteiinin mukana Western blot -analyysiin. Puhdistetun proteiinin k\u00e4ytt\u00e4minen positiivisena kontrollina on t\u00e4rke\u00e4\u00e4 tulosten varmistamiseksi, mutta t\u00e4ss\u00e4 tutkimuksessa sit\u00e4 ei ollut k\u00e4ytett\u00e4viss\u00e4.", "language": "fi", "element": "description", "qualifier": "abstract", "schema": "dc"}, {"key": "dc.description.abstract", "value": "Paajala, J. 2017. Immunoprecipitation of sirt1, sirt3 and PGC-1\u03b1 from rat gastrocnemius and soleus with magnetic beads. The Faculty of Sport and Health Sciences, University of Jyv\u00e4skyl\u00e4, Master\u2019s Thesis, 70 pp. \r\n\r\nSirt1, sirt3 and PGC-1\u03b1 are proteins that regulate among other factors the function and number of mitochondria. Understanding the biological function of these proteins is important, because risk factors and symptoms related to metabolic disorders are associated with poorly regulated mitochondrial function. Enzymatic differences between healthy and unhealthy individuals have been studied by using high capacity running/low capacity running rats contrasting model system. \r\n\r\nThe aim of the study was establish a method to isolate sirt1, sirt3 and PGC-1\u03b1 from gastrocnemius and soleus muscle samples of high capacity running rat by immunoprecipitation using magnetic beads. In addition, GAPDH and tubulin were investigated to validate the method further. The antibodies were incubated with the magnetic beads followed by incubation with homogenized muscle samples. Alternatively, antibodies and homogenized samples were incubated first followed by incubation with the beads. Next day, the beads were collected and the target proteins were eluted from the beads. The target proteins were analyzed using Western blot. \r\n\r\nThe results showed that sirt1, sirt3, PGC-1\u03b1, tubulin and GAPDH could not be isolated and \r\ndetected with the protocols described in this study. One issue was that antibodies used in immunoprecipitation co-eluted to the Western blot analysis interfering the imaging of the target protein. Second issue was the failure of the antibodies to bind to their target proteins or that antibodies did not attach to the magnetic beads and therefore some target proteins were washed away during immunoprecipitation. In Western blot, PGC-1\u03b1 was not detected at all. \r\n\r\nBy optimizing the conditions of the protocol, such as pH of buffers or changing antibodies, one might be able to isolate the target proteins. In addition, by using a crosslinking component between the antibody and the beads, co-elution of antibody to the Western blot analysis might possibly be avoided. To verify the results, the use of purified protein as a positive control is important, but in this study it was not available.", "language": "en", "element": "description", "qualifier": "abstract", "schema": "dc"}, {"key": "dc.description.provenance", "value": "Submitted using Plone Publishing form by Janne Paajala (japepaaj) on 2017-06-15 09:47:25.420953. Form: Pro gradu -lomake (https://kirjasto.jyu.fi/julkaisut/julkaisulomakkeet/pro-gradu-lomake). JyX data: [jyx_publishing-allowed (fi) =False]", "language": "en", "element": "description", "qualifier": "provenance", "schema": "dc"}, {"key": "dc.description.provenance", "value": "Submitted by jyx lomake-julkaisija (jyx-julkaisija.group@korppi.jyu.fi) on 2017-06-15T09:47:25Z\r\nNo. of bitstreams: 2\r\nURN:NBN:fi:jyu-201706152904.pdf: 1170448 bytes, checksum: de2417befadc823e671e2918fd3bee99 (MD5)\r\nlicense.html: 1170 bytes, checksum: 5ff8bd8407df231ee4e12a9d90a3f4bc (MD5)", "language": "en", "element": "description", "qualifier": "provenance", "schema": "dc"}, {"key": "dc.description.provenance", "value": "Made available in DSpace on 2017-06-15T09:47:25Z (GMT). No. of bitstreams: 2\r\nURN:NBN:fi:jyu-201706152904.pdf: 1170448 bytes, checksum: de2417befadc823e671e2918fd3bee99 (MD5)\r\nlicense.html: 1170 bytes, checksum: 5ff8bd8407df231ee4e12a9d90a3f4bc (MD5)\r\n Previous issue date: 2017", "language": "en", "element": "description", "qualifier": "provenance", "schema": "dc"}, {"key": "dc.format.extent", "value": "1 verkkoaineisto (77 sivua)", "language": null, "element": "format", "qualifier": "extent", "schema": "dc"}, {"key": "dc.format.mimetype", "value": "application/pdf", "language": null, "element": "format", "qualifier": "mimetype", "schema": "dc"}, {"key": "dc.language.iso", "value": "eng", "language": null, "element": "language", "qualifier": "iso", "schema": "dc"}, {"key": "dc.rights", "value": "In Copyright", "language": "en", "element": "rights", "qualifier": null, "schema": "dc"}, {"key": "dc.subject.other", "value": "immunoprecipitation", "language": null, "element": "subject", "qualifier": "other", "schema": "dc"}, {"key": "dc.subject.other", "value": "Western blot", "language": null, "element": "subject", "qualifier": "other", "schema": "dc"}, {"key": "dc.title", "value": "Immunoprecipitation of sirt1, sirt3 and PGC-1A from rat gastrocnemius and soleus with magnetic beads", "language": null, "element": "title", "qualifier": null, "schema": "dc"}, {"key": "dc.type", "value": "master thesis", "language": null, "element": "type", "qualifier": null, "schema": "dc"}, {"key": "dc.identifier.urn", "value": "URN:NBN:fi:jyu-201706152904", "language": null, "element": "identifier", "qualifier": "urn", "schema": "dc"}, {"key": "dc.type.ontasot", "value": "Pro gradu -tutkielma", "language": "fi", "element": "type", "qualifier": "ontasot", "schema": "dc"}, {"key": "dc.type.ontasot", "value": "Master\u2019s thesis", "language": "en", "element": "type", "qualifier": "ontasot", "schema": "dc"}, {"key": "dc.contributor.faculty", "value": "Liikuntatieteellinen tiedekunta", "language": "fi", "element": "contributor", "qualifier": "faculty", "schema": "dc"}, {"key": "dc.contributor.faculty", "value": "Faculty of Sport and Health Sciences", "language": "en", "element": "contributor", "qualifier": "faculty", "schema": "dc"}, {"key": "dc.contributor.department", "value": "Liikunta- ja terveystieteet", "language": "fi", "element": "contributor", "qualifier": "department", "schema": "dc"}, {"key": "dc.contributor.organization", "value": "University of Jyv\u00e4skyl\u00e4", "language": "en", "element": "contributor", "qualifier": "organization", "schema": "dc"}, {"key": "dc.contributor.organization", "value": "Jyv\u00e4skyl\u00e4n yliopisto", "language": "fi", "element": "contributor", "qualifier": "organization", "schema": "dc"}, {"key": "dc.subject.discipline", "value": "Liikuntafysiologia", "language": "fi", "element": "subject", "qualifier": "discipline", "schema": "dc"}, {"key": "dc.subject.discipline", "value": "Exercise Physiology", "language": "en", "element": "subject", "qualifier": "discipline", "schema": "dc"}, {"key": "dc.date.updated", "value": "2017-06-15T09:47:26Z", "language": "", "element": "date", "qualifier": "updated", "schema": "dc"}, {"key": "yvv.contractresearch.funding", "value": "0", "language": "", "element": "contractresearch", "qualifier": "funding", "schema": "yvv"}, {"key": "dc.type.coar", "value": "http://purl.org/coar/resource_type/c_bdcc", "language": null, "element": "type", "qualifier": "coar", "schema": "dc"}, {"key": "dc.rights.accesslevel", "value": "restrictedAccess", "language": "fi", "element": "rights", "qualifier": "accesslevel", "schema": "dc"}, {"key": "dc.type.publication", "value": "masterThesis", "language": null, "element": "type", "qualifier": "publication", "schema": "dc"}, {"key": "dc.subject.oppiainekoodi", "value": "5011", "language": null, "element": "subject", "qualifier": "oppiainekoodi", "schema": "dc"}, {"key": "dc.subject.yso", "value": "sirtuiinit", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.format.content", "value": "fulltext", "language": null, "element": "format", "qualifier": "content", "schema": "dc"}, {"key": "dc.rights.url", "value": "https://rightsstatements.org/page/InC/1.0/", "language": null, "element": "rights", "qualifier": "url", "schema": "dc"}, {"key": "dc.rights.accessrights", "value": "Aineistoon p\u00e4\u00e4sy\u00e4 on rajoitettu tekij\u00e4noikeussyist\u00e4. Aineisto on luettavissa Jyv\u00e4skyl\u00e4n yliopiston kirjaston arkistoty\u00f6asemalta. Ks. https://kirjasto.jyu.fi/fi/tyoskentelytilat/laitteet-ja-tilat.", "language": "fi", "element": "rights", "qualifier": "accessrights", "schema": "dc"}, {"key": "dc.rights.accessrights", "value": "This material has a restricted access due to copyright reasons. It can be read at the workstation at Jyv\u00e4skyl\u00e4 University Library reserved for the use of archival materials: https://kirjasto.jyu.fi/en/workspaces/facilities.", "language": "en", "element": "rights", "qualifier": "accessrights", "schema": "dc"}, {"key": "dc.type.okm", "value": "G2", "language": null, "element": "type", "qualifier": "okm", "schema": "dc"}]
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