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.|
Sensitive: The biotin tag allows detection of 0.5-5ng of translated protein.Safe: No radioisotope handling, storage or disposal is required.Fast: Labeled proteins can be detected 3-4 hours after gel electrophoresis.Flexible: Results can be visualized by using colorimetric or chemiluminescent detection.