The Availability of a Recombinant Anti-SNAP Antibody in VHH Format Amplifies the Application Flexibility of SNAP-Tagged Proteins

المؤلفون المشاركون

de Marco, Ario
Aliprandi, Marisa
Pivetta, Flavia
Maestro, Roberta
Sparacio, Eleonora
Ossolengo, Giuseppe

المصدر

Journal of Biomedicine and Biotechnology

العدد

المجلد 2010، العدد 2010 (31 ديسمبر/كانون الأول 2010)، ص ص. 1-7، 7ص.

الناشر

Hindawi Publishing Corporation

تاريخ النشر

2010-10-17

دولة النشر

مصر

عدد الصفحات

7

التخصصات الرئيسية

الطب البشري

الملخص 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.

نمط استشهاد جمعية علماء النفس الأمريكية (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

نمط استشهاد الجمعية الأمريكية للغات الحديثة (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

نمط استشهاد الجمعية الطبية الأمريكية (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

نوع البيانات

مقالات

لغة النص

الإنجليزية

الملاحظات

Includes bibliographical references

رقم السجل

BIM-488950