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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.
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.