| Autor: |
Yuri Emoto, Ryoya Katayama, Emi Hibino, Sho Ishihara, Natsuko Goda, Takeshi Tenno, Yoshihiro Kobashigawa, Hiroshi Morioka, Hidekazu Hiroaki |
| Jazyk: |
angličtina |
| Rok vydání: |
2023 |
| Předmět: |
|
| Zdroj: |
Methods and Protocols, Vol 6, Iss 3, p 44 (2023) |
| Druh dokumentu: |
article |
| ISSN: |
2409-9279 |
| DOI: |
10.3390/mps6030044 |
| Popis: |
The detection and quantification of protein–protein interactions (PPIs) is a crucial technique that often involves the use of recombinant proteins with fusion protein tags, such as maltose-binding protein (MBP) and glutathione-S-transferase (GST). In this study, we improved the cohesive and sticky properties of gelatinized starch by supplementing it with agarose, resulting in a harder gel that could coat the bottom of a microtiter plate. The resulting gelatinized starch/agarose mixture allowed for the efficient immobilization of MBP-tagged proteins on the coated plates, enabling the use of indirect ELISA-like PPI assays. By using the enzymatic activity of GST as an indicator, we succeeded in determining the dissociation constants between MBP-tagged and GST-tagged proteins on 96-well microtiter plates and a microplate reader without any expensive specialized equipment. |
| Databáze: |
Directory of Open Access Journals |
| Externí odkaz: |
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