Specific Protein Quantification by Radioimmuno-Dot-Blot Assay for Complex Mixture Samples Utilizing Strep-Tag and Tritium-Labeled Strep-Tactin.

ABSTRACT: Accurately quantifying specific proteins from complex mixtures like cell lysates, for example, during in vivo studies, is difficult, especially for aggregation-prone proteins. Herein, we describe the development of a specific protein quantification method that combines a solid-state dot blot approach with radiolabel detection via liquid scintillation counting. The specific detection with high sensitivity is achieved by using the Twin-Strep protein affinity tag and tritium-labeled ^3HStrep-TactinXT probe. While the assay was developed with the recombinant silk protein CBM-AQ12-CBM as a target, the method can be adapted to other recombinant proteins. Variations of the protein tag and Strep-Tactin probe were tested, and it was found that only the combination of Strep-TactinXT and Tw

SUBMITTER: Malkamaki M 

PROVIDER: S-EPMC11755393 | biostudies-literature | 2025 Jan

REPOSITORIES: biostudies-literature