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The Availability of a Recombinant Anti-SNAP Antibody in VHH Format Amplifies the Application Flexibility of SNAP-Tagged Proteins
Joint Authors
de Marco, Ario
Aliprandi, Marisa
Pivetta, Flavia
Maestro, Roberta
Sparacio, Eleonora
Ossolengo, Giuseppe
Source
Journal of Biomedicine and Biotechnology
Issue
Vol. 2010, Issue 2010 (31 Dec. 2010), pp.1-7, 7 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2010-10-17
Country of Publication
Egypt
No. of Pages
7
Main Subjects
Abstract EN
Antibodies are indispensable reagents in basic research, and those raised against tags constitute a useful tool for the evaluation of the biochemistry and biology of novel proteins.
In this paper, we describe the isolation and characterization of a single-domain recombinant antibody (VHH) specific for the SNAP-tag, using Twist2 as a test-protein.
The antibody was efficient in western blot, immunoprecipitation, immunopurification, and immunofluorescence.
The sequence corresponding to the anti-SNAP has been subcloned for large-scale expression in vectors that allow its fusion to either a 6xHis-tag or the Fc domain of rabbit IgG2 taking advantage of a new plasmid that was specifically designed for VHH antibodies.
The two different fusion antibodies were compared in immunopurification and immunofluorescence experiments, and the recombinant protein SNAP-Twist2 was accurately identified by the anti-SNAP Fc-VHH construct in the nuclear/nucleolar subcellular compartment.
Furthermore, such localization was confirmed by direct Twist2 identification by means of anti-Twisit2 VHH antibodies recovered after panning of the same naïve phage display library used to isolate the anti-SNAP binders.
Our successful localization of Twist2 protein using the SNAP-tag-based approach and the anti-Twist2-specific recombinant single-domain antibodies opens new research possibilities in this field.
American Psychological Association (APA)
Aliprandi, Marisa& Sparacio, Eleonora& Pivetta, Flavia& Ossolengo, Giuseppe& Maestro, Roberta& de Marco, Ario. 2010. The Availability of a Recombinant Anti-SNAP Antibody in VHH Format Amplifies the Application Flexibility of SNAP-Tagged Proteins. Journal of Biomedicine and Biotechnology،Vol. 2010, no. 2010, pp.1-7.
https://search.emarefa.net/detail/BIM-488950
Modern Language Association (MLA)
Aliprandi, Marisa…[et al.]. The Availability of a Recombinant Anti-SNAP Antibody in VHH Format Amplifies the Application Flexibility of SNAP-Tagged Proteins. Journal of Biomedicine and Biotechnology No. 2010 (2010), pp.1-7.
https://search.emarefa.net/detail/BIM-488950
American Medical Association (AMA)
Aliprandi, Marisa& Sparacio, Eleonora& Pivetta, Flavia& Ossolengo, Giuseppe& Maestro, Roberta& de Marco, Ario. The Availability of a Recombinant Anti-SNAP Antibody in VHH Format Amplifies the Application Flexibility of SNAP-Tagged Proteins. Journal of Biomedicine and Biotechnology. 2010. Vol. 2010, no. 2010, pp.1-7.
https://search.emarefa.net/detail/BIM-488950
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-488950