Can I detect a protein expressed in the TNT® Coupled Transcription/Translation using Coomassie® blue-stained SDS-PAGE?
No, your protein cannot be seen on a Coomassie® blue-stained SDS polyacrylamide gel (SDS-PAGE) if translated using the TNT® Coupled Transcription/Translation System. Since the in vitro protein expression system itself is a pool of proteins of various sizes, it is impossible to distinguish nanogram amounts of newly synthesized protein from the background of other proteins. To detect a single protein, one commonly used method is to add a labeled amino acid (e.g., [35S]Methionine) to the TNT® reaction to be incorporated into the synthesized protein followed by detection using SDS-PAGE and autoradiography. Alternatively, non-radioactive protein-labeling methods can be used such as the Transcend™ tRNA or FluoroTect™ GreenLys in vitro Translation Labeling System. With the Transcend™ Non-Radioactive Labeling System, biotinylated lysine is incorporated into the newly synthesized proteins and detected by colorimetric (Cat.# L5070) or chemiluminescent (Cat.# L5080) methods after Western blotting
Related Questions
- Can I detect a protein expressed in the TNT® Coupled Transcription/Translation using Coomassie® blue-stained SDS-PAGE?
- Are transcription (RNA synthesis) or translation (protein synthesis) required for cleavage and/or early development?
- What is the difference between transcription, and translation in protein synthesis
