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dc.contributor.author
Brun, Antonio
dc.contributor.author
Magallanes Alba, Melisa Eliana
dc.contributor.author
Martínez del Rio, Carlos
dc.contributor.author
Barrett Wilt, Gregory
dc.contributor.author
Karasov, William H.
dc.contributor.author
Caviedes Vidal, Enrique Juan Raul
dc.date.available
2021-10-26T10:27:01Z
dc.date.issued
2020-02
dc.identifier.citation
Brun, Antonio; Magallanes Alba, Melisa Eliana; Martínez del Rio, Carlos; Barrett Wilt, Gregory; Karasov, William H.; et al.; A fast and accurate method to identify and quantify enzymes in brush-border membranes: In situ hydrolysis followed by nano LC-MS/MS; Molecular Diversity Preservation International; Methods and Protocols; 3; 1; 2-2020; 1-9
dc.identifier.issn
2409-9279
dc.identifier.uri
http://hdl.handle.net/11336/145000
dc.description.abstract
A simple method for the identification of brush-border membrane α-glucosidases is described. The proteins were first solubilized and separated in a gel under native, non-denaturing, conditions. The gel was then incubated in substrate solutions (maltose or sucrose), and the product (glucose) exposed in situ by the oxidation of o-dianisidine, which yields a brown-orange color. Nano-liquid chromatography coupled to mass spectrometry analyses of proteins (nano LC-MS/MS) present in the colored bands excised from the gels, was used to confirm the presence of the enzymes. The stain is inexpensive and the procedure permits testing several substrates in the same gel. Once enzymes are identified, their abundance, relative to that of other proteins in the brush border, can be semi-quantified using nano LC-MS/MS.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
Molecular Diversity Preservation International
dc.rights
info:eu-repo/semantics/openAccess
dc.rights.uri
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.subject
α-glucosidase
dc.subject
polyacrylamide gels
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proteomics
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activity
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detection
dc.subject.classification
Métodos de Investigación en Bioquímica
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Ciencias Biológicas
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CIENCIAS NATURALES Y EXACTAS
dc.title
A fast and accurate method to identify and quantify enzymes in brush-border membranes: In situ hydrolysis followed by nano LC-MS/MS
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2021-09-07T13:56:43Z
dc.journal.volume
3
dc.journal.number
1
dc.journal.pagination
1-9
dc.journal.pais
Suiza
dc.description.fil
Fil: Brun, Antonio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis. Universidad Nacional de San Luis. Facultad de Ciencias Físico Matemáticas y Naturales. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis; Argentina
dc.description.fil
Fil: Magallanes Alba, Melisa Eliana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis. Universidad Nacional de San Luis. Facultad de Ciencias Físico Matemáticas y Naturales. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis; Argentina
dc.description.fil
Fil: Martínez del Rio, Carlos. University of Wyoming; Estados Unidos
dc.description.fil
Fil: Barrett Wilt, Gregory. University of Wisconsin; Estados Unidos
dc.description.fil
Fil: Karasov, William H.. University of Wisconsin; Estados Unidos
dc.description.fil
Fil: Caviedes Vidal, Enrique Juan Raul. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis. Universidad Nacional de San Luis. Facultad de Ciencias Físico Matemáticas y Naturales. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis; Argentina
dc.journal.title
Methods and Protocols
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/https://www.mdpi.com/2409-9279/3/1/15
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.3390/mps3010015
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