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
Issue
Vol. 2010, Issue 2010 (31 Dec. 2009), pp.1-7, 7 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2010-09-26
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. BioMed Research International،Vol. 2010, no. 2010, pp.1-7.
https://search.emarefa.net/detail/BIM-989056
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. BioMed Research International No. 2010 (Dec. 2010), pp.1-7.
https://search.emarefa.net/detail/BIM-989056
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. BioMed Research International. 2010. Vol. 2010, no. 2010, pp.1-7.
https://search.emarefa.net/detail/BIM-989056
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-989056