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Abstract for the Transcend™ Non-Radioactive Translation Detection Systems
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, ε-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, 0.5–5ng of protein can be
detected by these methods within 3–4 hours after gel electrophoresis.
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