Comparision of protein precipitation methods for two-dimensional electrophoresis of dog salivary proteins
Autor(a) principal: | |
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Data de Publicação: | 2018 |
Outros Autores: | , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10174/24357 https://doi.org/10.5584/jiomics.v8i1.232 |
Resumo: | Despite saliva being one fluid with growing interest as a source of biomarkers, both in humans and animal models, few studies have been reported that use proteomic approaches for canine saliva analyses. Two-dimensional electrophoresis (2-DE) is considerably used in biomarker research and its use for dog saliva study may had relevant knowledge about pathology/physiology. The quality of the results obtained using 2-DE greatly depends on sample preparation. Different protein precipitation methods are frequently used for removing interfering compounds and concentrating samples, but their efficiency varies according to sample characteristics. For dog saliva samples no information was found about the best precipitant and precipitation method for electrophoretic protein profiling. In this study, six different protein precipitation methods were compared. Precipitation of dog salivary proteins with 20% (w/v) trichloroacetic acid (TCA) resulted in lower protein recovery rate than other methods tested, but allowed protein profiles highly correlated with the ones from original samples. Moreover, this protocol resulted in good protein separation in 2-DE, with the visualization of spots from salivary proteins not observed when samples were treated using other methods. Based on this, we propose the use of TCA for dog saliva whenever precipitation is needed for protein profile analysis. |
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Comparision of protein precipitation methods for two-dimensional electrophoresis of dog salivary proteinsdogsalivaprotein precipitationtwo-dimensional electrophoresisDespite saliva being one fluid with growing interest as a source of biomarkers, both in humans and animal models, few studies have been reported that use proteomic approaches for canine saliva analyses. Two-dimensional electrophoresis (2-DE) is considerably used in biomarker research and its use for dog saliva study may had relevant knowledge about pathology/physiology. The quality of the results obtained using 2-DE greatly depends on sample preparation. Different protein precipitation methods are frequently used for removing interfering compounds and concentrating samples, but their efficiency varies according to sample characteristics. For dog saliva samples no information was found about the best precipitant and precipitation method for electrophoretic protein profiling. In this study, six different protein precipitation methods were compared. Precipitation of dog salivary proteins with 20% (w/v) trichloroacetic acid (TCA) resulted in lower protein recovery rate than other methods tested, but allowed protein profiles highly correlated with the ones from original samples. Moreover, this protocol resulted in good protein separation in 2-DE, with the visualization of spots from salivary proteins not observed when samples were treated using other methods. Based on this, we propose the use of TCA for dog saliva whenever precipitation is needed for protein profile analysis.Journal of Integrated Omics2019-01-30T11:23:34Z2019-01-302018-04-18T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttp://hdl.handle.net/10174/24357http://hdl.handle.net/10174/24357https://doi.org/10.5584/jiomics.v8i1.232englucenazen@gmail.comndndfcs@uevora.ptasta@um.esecsl@uevora.pt205Lucena, SóniaCarreira, MariaRodrigues, LéniaCapela e Silva, FernandoTvarijonaviciute, AstaLamy, Elsainfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-01-03T19:17:21Zoai:dspace.uevora.pt:10174/24357Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T01:15:03.988474Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
Comparision of protein precipitation methods for two-dimensional electrophoresis of dog salivary proteins |
title |
Comparision of protein precipitation methods for two-dimensional electrophoresis of dog salivary proteins |
spellingShingle |
Comparision of protein precipitation methods for two-dimensional electrophoresis of dog salivary proteins Lucena, Sónia dog saliva protein precipitation two-dimensional electrophoresis |
title_short |
Comparision of protein precipitation methods for two-dimensional electrophoresis of dog salivary proteins |
title_full |
Comparision of protein precipitation methods for two-dimensional electrophoresis of dog salivary proteins |
title_fullStr |
Comparision of protein precipitation methods for two-dimensional electrophoresis of dog salivary proteins |
title_full_unstemmed |
Comparision of protein precipitation methods for two-dimensional electrophoresis of dog salivary proteins |
title_sort |
Comparision of protein precipitation methods for two-dimensional electrophoresis of dog salivary proteins |
author |
Lucena, Sónia |
author_facet |
Lucena, Sónia Carreira, Maria Rodrigues, Lénia Capela e Silva, Fernando Tvarijonaviciute, Asta Lamy, Elsa |
author_role |
author |
author2 |
Carreira, Maria Rodrigues, Lénia Capela e Silva, Fernando Tvarijonaviciute, Asta Lamy, Elsa |
author2_role |
author author author author author |
dc.contributor.author.fl_str_mv |
Lucena, Sónia Carreira, Maria Rodrigues, Lénia Capela e Silva, Fernando Tvarijonaviciute, Asta Lamy, Elsa |
dc.subject.por.fl_str_mv |
dog saliva protein precipitation two-dimensional electrophoresis |
topic |
dog saliva protein precipitation two-dimensional electrophoresis |
description |
Despite saliva being one fluid with growing interest as a source of biomarkers, both in humans and animal models, few studies have been reported that use proteomic approaches for canine saliva analyses. Two-dimensional electrophoresis (2-DE) is considerably used in biomarker research and its use for dog saliva study may had relevant knowledge about pathology/physiology. The quality of the results obtained using 2-DE greatly depends on sample preparation. Different protein precipitation methods are frequently used for removing interfering compounds and concentrating samples, but their efficiency varies according to sample characteristics. For dog saliva samples no information was found about the best precipitant and precipitation method for electrophoretic protein profiling. In this study, six different protein precipitation methods were compared. Precipitation of dog salivary proteins with 20% (w/v) trichloroacetic acid (TCA) resulted in lower protein recovery rate than other methods tested, but allowed protein profiles highly correlated with the ones from original samples. Moreover, this protocol resulted in good protein separation in 2-DE, with the visualization of spots from salivary proteins not observed when samples were treated using other methods. Based on this, we propose the use of TCA for dog saliva whenever precipitation is needed for protein profile analysis. |
publishDate |
2018 |
dc.date.none.fl_str_mv |
2018-04-18T00:00:00Z 2019-01-30T11:23:34Z 2019-01-30 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10174/24357 http://hdl.handle.net/10174/24357 https://doi.org/10.5584/jiomics.v8i1.232 |
url |
http://hdl.handle.net/10174/24357 https://doi.org/10.5584/jiomics.v8i1.232 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
lucenazen@gmail.com nd nd fcs@uevora.pt asta@um.es ecsl@uevora.pt 205 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.publisher.none.fl_str_mv |
Journal of Integrated Omics |
publisher.none.fl_str_mv |
Journal of Integrated Omics |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
instname_str |
Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
instacron_str |
RCAAP |
institution |
RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
collection |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository.name.fl_str_mv |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
repository.mail.fl_str_mv |
|
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1799136632683626496 |