Protocol for the production and purification of an i-Motif-specific nanobody

Autor: Mahdi Zeraati, Samuel E. Ross, Behnaz Aghaei, Alvaro González Rajal, Cecile King, Marcel E. Dinger
Jazyk: angličtina
Rok vydání: 2023
Předmět:
Zdroj: STAR Protocols, Vol 4, Iss 4, Pp 102729- (2023)
Druh dokumentu: article
ISSN: 2666-1667
DOI: 10.1016/j.xpro.2023.102729
Popis: Summary: Intercalated motifs or i-Motifs (iMs) are nucleic acid structures formed by cytosine-rich sequences, which may regulate cellular processes and have broad applications in nanotechnology due to their pH-dependent nature. We have developed an iM-specific nanobody (iMbody) that can recognize iM DNA structures regardless of their sequences, making it a versatile research tool for studying iMs in various contexts. Here, we provide a protocol for the bacterial expression and His-tag purification of iMbody. We then describe procedures for performing ELISA and immunostaining using iMbody. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
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