Fast-dissociating but highly specific antibodies are novel tools in biology, especially useful for multiplex super-resolution microscopy

Summary: Fast-dissociating, highly specific monoclonal antibodies (FDSAs) are single-molecule imaging probes useful for many biological assays including consecutive, multiplexable super-resolution microscopy. We developed a screening assay to characterize the kinetics of antibody-antigen interaction...

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Bibliographic Details
Published in:STAR Protocols
Main Authors: Takushi Miyoshi, Thomas B. Friedman, Naoki Watanabe
Format: Article
Language:English
Published: Elsevier 2021-12-01
Subjects:
Single-molecule Assays
High Throughput Screening
Microscopy
Antibody
Molecular/Chemical Probes
Protein Biochemistry
Online Access:http://www.sciencedirect.com/science/article/pii/S2666166721006730
Description
Summary:Summary: Fast-dissociating, highly specific monoclonal antibodies (FDSAs) are single-molecule imaging probes useful for many biological assays including consecutive, multiplexable super-resolution microscopy. We developed a screening assay to characterize the kinetics of antibody-antigen interactions using single-molecule microscopy and established a pipeline to identify FDSAs from thousands of monoclonal candidates. Provided here are detailed protocols to prepare multi-well glass-bottom plates necessary for our assay to identify hybridoma clones secreting FDSAs. Synthesis of fluorescently labeled Fab fragments (Fab probes) from FDSAs is also described.For complete details on the use and execution of this protocol, please refer to Miyoshi et al. (2021).
ISSN:2666-1667

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