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 Phosph...

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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, 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.

Features - Benefits

• 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.

Applications

For more information, see the Protocols & Applications Guide.

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