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dc.contributor.author
Zalazar, Lucia
dc.contributor.author
Alonso, Carlos Agustín Isidro
dc.contributor.author
de Castro, Rosana Esther
dc.contributor.author
Cesari, Andreina
dc.date.available
2017-02-22T14:01:20Z
dc.date.issued
2014-01
dc.identifier.citation
Zalazar, Lucia; Alonso, Carlos Agustín Isidro; de Castro, Rosana Esther; Cesari, Andreina; An alternative easy method for antibody purification and analysis of protein-protein interaction-using GST fusion proteins immobilized onto glutathione-agarose; Springer; Analytical And Bioanalytical Chemistry; 406; 3; 1-2014; 911-914
dc.identifier.issn
1618-2642
dc.identifier.uri
http://hdl.handle.net/11336/13285
dc.description.abstract
Immobilization of small proteins designed to perform protein-protein assays can be a difficult task. Often, the modification of reactive residues necessary for the interaction between the immobilized protein and the matrix compromise the interaction between the protein and its target. In these cases, GST is a valuable tag providing a long arm that makes the bait protein to be accessible to the mobile flow phase of the chromatography. In the present report, we used a GST-fusion version of the 8 kDa protein SPINK3 as the bait to purify anti-SPINK3 antibodies from a rabbit crude serum. The protocol for immobilization of GST-SPINK3 to glutathione agarose beads was modified from previously reported protocols by using an alternative bifunctional crosslinker (DSP) in a very simple procedure and by using simple buffers under physiological conditions. We concluded that the immobilized protein remained bound to the column after elution with low pH, allowing the re-use of the column for alternative uses, such as screening for other protein-protein interactions using SPINK3 as bait.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
Springer
dc.rights
info:eu-repo/semantics/restrictedAccess
dc.rights.uri
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.subject
Antibody Purification
dc.subject
Protein-Protein Interactions
dc.subject
Glutathione-Agarose Column
dc.subject
Dsp Cross-Linker
dc.subject
Spink3
dc.subject.classification
Biología Reproductiva
dc.subject.classification
Ciencias Biológicas
dc.subject.classification
CIENCIAS NATURALES Y EXACTAS
dc.title
An alternative easy method for antibody purification and analysis of protein-protein interaction-using GST fusion proteins immobilized onto glutathione-agarose
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2017-02-21T14:37:02Z
dc.identifier.eissn
1618-2650
dc.journal.volume
406
dc.journal.number
3
dc.journal.pagination
911-914
dc.journal.pais
Alemania
dc.journal.ciudad
Berlín
dc.description.fil
Fil: Zalazar, Lucia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Mar del Plata. Instituto de Investigaciones Biológicas; Argentina
dc.description.fil
Fil: Alonso, Carlos Agustín Isidro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Mar del Plata. Instituto de Investigaciones Biológicas; Argentina
dc.description.fil
Fil: de Castro, Rosana Esther. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Mar del Plata. Instituto de Investigaciones Biológicas; Argentina
dc.description.fil
Fil: Cesari, Andreina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Mar del Plata. Instituto de Investigaciones Biológicas; Argentina
dc.journal.title
Analytical And Bioanalytical Chemistry
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/http://link.springer.com/article/10.1007%2Fs00216-013-7533-6
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.1007/s00216-013-7533-6


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