The TNT® Quick Systems are convenient single-tube, coupled transcription/translation reactions for eukaryotic cell-free protein expression. These cell-free expression systems combine the RNA Polymerase, nucleotides, salts, amino acids and Recombinant RNasin® Ribonuclease Inhibitor with the reticulocyte lysate solution to form a single TNT® Quick Master Mix. The TNT® Quick Coupled Transcription/Translation System is available in two configurations for the expression of genes cloned downstream from either the T7 or SP6 RNA polymerase promoters. To use these cell-free expression systems, 0.2–2.0μg of circular plasmid DNA containing a T7 or SP6 promoter, or a PCRgenerated fragment containing a T7 promoter, is added to an aliquot of the TNT® Quick Master Mix and incubated in a 50μl reaction volume for 60–90 minutes at 30°C. The expression reaction produces significant quantities of protein for a variety of applications including GST pull-downs and gel shift assays.

Applications (of cell-free expression)

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

For more information about cell-free protein expression, see the Promega Protocols & Applications Guide at www.promega.com/paguide.

Features:

• Use in Multiple Applications: The TNT® cell-free expression systems are widely used for protein:protein interaction, protein:nucleic acid interactions, and more.
• Save Time: Using a one-tube reaction, proteins are expressed 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 a cell-free mammalian system.
• Dependability You Can Count On: The TNT® Systems are rigorously quality controlled to ensure the highest level of protein expression.

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