TnT® Coupled Reticulocyte Lysate Systems

The TnT® Coupled Reticulocyte Lysate Systems offer researchers an alternative for eukaryotic cell-free protein expression: a single-tube, coupled transcription/translation system. The TnT® Lysate Systems greatly simplify the process and reduce the time required to obtain in vitro translation results. Standard rabbit reticulocyte lysate translations commonly use RNA synthesized in vitro from SP6, T3 or T7 RNA polymerase promoters and require three separate reactions with several steps between each reaction. The TnT® Systems bypass many of these steps by incorporating transcription directly in the translatio...

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The TnT® Coupled Reticulocyte Lysate Systems offer researchers an alternative for eukaryotic cell-free protein expression: a single-tube, coupled transcription/translation system. The TnT® Lysate Systems greatly simplify the process and reduce the time required to obtain in vitro translation results. Standard rabbit reticulocyte lysate translations commonly use RNA synthesized in vitro from SP6, T3 or T7 RNA polymerase promoters and require three separate reactions with several steps between each reaction. The TnT® Systems bypass many of these steps by incorporating transcription directly in the translation mix.

For optimal protein expression using the TnT® SP6 RNA polymerase, we recommend titrating magnesium acetate in 0.1mM increments between 0.1mM and 0.5mM. In some instances the addition of 0.2mM magnesium acetate has been shown to increase protein expression by 40%. Magnesium acetate is supplied only with Cat.# L4600 and L4601.

Features - Benefits

• Use in Multiple Applications: The TnT® Systems are widely used for protein:protein interaction, protein:nucleic acid interactions, and more.
• Save Time: Using a one-tube reaction, proteins are generated in one hour, not days, as with in vivo methods.
• Complete System: All the reagents you need are provided (except radioisotopes).
• Reliable: Eliminate solubility issues by using an in vitro mammalian system.
• Dependability You Can Count On: The TnT® Systems are rigorously quality controlled to ensure the highest level of performance.

Applications

• GST pull-downs (view an animation).
• Gel shift assays (view an animation).
• Co-immunoprecipitation (view an animation).
• Characterization of protein modifications (view an animation).
• Protein:RNA interactions.
• Protein activity studies.
• Confirmation of gene products.
• Protein Truncation Test (PTT).

For more information about cell-free protein expression, see the Protein Expression chapter of the Protocols & Applications Guide.

References

1. Pelham, H.R.B. and Jackson, R.J. (1976) Eur. J. Biochem. 67, 247–56.
2. Krieg, P.A. and Melton, D.A. (1984) Nucl. Acids Res. 12, 7057–70.

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