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ファイル | 記述 | サイズ | フォーマット | |
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j.xpro.2023.102523.pdf | 5.75 MB | Adobe PDF | 見る/開く |
タイトル: | Protocol to generate fast-dissociating recombinant antibody fragments for multiplexed super-resolution microscopy |
著者: | Zhang, Qianli Miyamoto, Akitoshi Watanabe, Naoki https://orcid.org/0000-0001-8492-200X (unconfirmed) |
著者名の別形: | 張, 千里 宮本, 章歳 渡邊, 直樹 |
発行日: | 15-Sep-2023 |
出版者: | Elsevier BV |
誌名: | STAR Protocols |
巻: | 4 |
号: | 3 |
論文番号: | 102523 |
抄録: | Multiplexed high-density label super-resolution microscopy image reconstruction by integrating exchangeable single-molecule localization (IRIS) enables elucidating fine structures and molecular distribution in cells and tissues. However, fast-dissociating binders are required for individual targets. Here, we present a protocol for generating antibody-based IRIS probes from existing antibody sequences. We describe steps for retrieving antibody sequences from databases. We then detail the construction, purification, and evaluation of recombinant probes after site-directed mutagenesis at the base of complementarity-determining region loops. The protocol accelerates dissociation rates without compromising the binding specificity. For complete details on the use and execution of this protocol, please refer to Zhang et al. (2022). |
著作権等: | © 2023 The Authors. This is an open access article under the CC BY-NC-ND license. |
URI: | http://hdl.handle.net/2433/286624 |
DOI(出版社版): | 10.1016/j.xpro.2023.102523 |
PubMed ID: | 37610875 |
出現コレクション: | 学術雑誌掲載論文等 |
このアイテムは次のライセンスが設定されています: クリエイティブ・コモンズ・ライセンス