| RRC ID |
72103
|
| Author |
Miyoshi T, Friedman TB, Watanabe N.
|
| Title |
Fast-dissociating but highly specific antibodies are novel tools in biology, especially useful for multiplex super-resolution microscopy.
|
| Journal |
STAR Protoc
|
| Abstract |
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).
|
| Volume |
2(4)
|
| Pages |
100967
|
| Published |
2021-12-17
|
| DOI |
10.1016/j.xpro.2021.100967
|
| PII |
S2666-1667(21)00673-0
|
| PMID |
34841279
|
| PMC |
PMC8605432
|
| MeSH |
Animals
Antibodies, Monoclonal* / chemistry
Antibodies, Monoclonal* / metabolism
Female
Fluorescent Dyes* / chemistry
Fluorescent Dyes* / metabolism
Hybridomas / chemistry
Hybridomas / metabolism
Immunoglobulin Fab Fragments / chemistry
Immunoglobulin Fab Fragments / metabolism
Mice
Mice, Inbred BALB C
Microscopy, Fluorescence / methods*
Single Molecule Imaging / methods*
|
| Resource |
| Human and Animal Cells |
UV-P3U1-B8(RCB1157) |
