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

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...

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Bibliographic Details
Main Authors: Mahdi Zeraati, Samuel E. Ross, Behnaz Aghaei, Alvaro González Rajal, Cecile King, Marcel E. Dinger
Format: Article
Language:English
Published: Elsevier 2023-12-01
Series:STAR Protocols
Subjects:
Online Access:http://www.sciencedirect.com/science/article/pii/S2666166723006962
Description
Summary: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.
ISSN:2666-1667