Non-Radioactive Translation Detection Systems for colorimetric or chemiluminescent detection of proteins synthesized in vitro. Biotinylated proteins are detected using Streptavidin-Alkaline Phosphatase or Streptavidin-HRP.
- Non-Radioactive Detection of In Vitro Translated Proteins
- Biotin tag allows detection of 0.5-5ng of protein 3-4 hours after gel electrophoresis
- No radioisotope handling, storage or disposal issues
- Option of colorimetric or chemiluminescent detection
The Transcend Non-Radioactive Translation Detection Systems allow non-radioactive detection of proteins synthesized in vitro. Using these systems, biotinylated lysine residues are incorporated into nascent proteins during translation, eliminating the need for labeling with [35S]methionine or other radioactive amino acids. This biotinylated lysine is added to the translation reaction as a precharged epsilon-labeled biotinylated lysine-tRNA complex (Transcend tRNA) rather than a free amino acid. After SDS-PAGE and electroblotting, the biotinylated proteins can be visualized by binding either Streptavidin-Alkaline Phosphatase (Streptavidin-AP) or Streptavidin-Horseradish Peroxidase (Streptavidin-HRP), followed either by colorimetric or chemiluminescent detection. Typically, these methods can detect 0.5-5ng of protein within 3-4 hours after gel electrophoresis. This sensitivity is equivalent to that achieved with [35S]methionine incorporation and autoradiographic detection 6-12 hours after gel electrophoresis.
After SDS-PAGE and electroblotting, users can visualize the biotinylated proteins by binding either Streptavidin-Alkaline Phosphatase (Streptavidin-AP) or Streptavidin-Horseradish Peroxidase (Streptavidin-HRP) followed either by colorimetric or chemiluminescent detection. Typically, these methods can detect 0.5 to 5 ng of protein within three-to-four hours after gel electrophoresis. This sensitivity is equivalent to that achieved with [35S]methionine incorporation and autoradiographic detection six to 12 hours after gel electrophoresis.
This biotinylated lysine is added to the translation reaction as a pre-charged ε-labeled biotinylated lysine-tRNA complex (Transcend tRNA) rather than a free amino acid.
