TnT® Quick Coupled Transcription/Translation System
Single-Tube, Coupled Transcription/Translation for Eukaryotic Cell-Free Protein Expression
- Functional protein is expressed in one hour, ready for use without further purification
- Use protein for protein:protein and protein:nucleic acid interactions, protein modification and more
- Choice of SP6 or T7 promoter
Catalog Number:
Choose a Promoter
Size
Catalog Number: L1170
Catalog Number: L1171
Catalog Number: L2080
Catalog Number: L2081
Express Protein from PCR Fragments or Plasmid DNA
The TnT® Quick Coupled Transcription/Translation Systems provide convenient single-tube, coupled transcription/translation reactions for eukaryotic cell-free protein expression. These in vitro translation 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 protein expression systems, 0.2–2.0μg of circular plasmid DNA containing a T7 or SP6 promoter, or a PCR-generated 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.
These mammalian-based systems express soluble, functional proteins that are post-translationally modified, unlike E. coli-based systems. Expressed proteins can be used directly after expression; no additional protein purification needed.
Schematic showing DNA to protein expression in 60–90 minutes.
Applications of In Vitro Translation Systems
The TnT® Transcription/Translation Systems provide a simplified and accelerated method to perform in vitro protein translation. These systems have proven to be useful for many downstream applications including, but not limited to, glutathione-S-transferase (GST) pull-downs, gel shift assays, co-immunoprecipitation, characterization of protein modifications, protein:protein and protein:nucleic acid interactions and protein activity studies.
The animations and article below discuss a few of these applications in more detail. For more information about in vitro protein translation systems and how they can be used, see the Protein Expression chapter of the Protocols & Applications Guide.
GST Pull-Downs
Gel Shift Assays
Co-immunoprecipitation
Characterization of Protein Modifications
Further Downstream: Expanded Applications for TnT® Transcription/Translation Systems
protein translation. These systems are useful for many downstream applications,
including protein:protein interactions and protein:DNA interactions.
In this article, we review several publications that utilized TnT® Systems and
explore how these systems can be employed beyond these initial protein
translation applications.
Protocols
Specifications
Catalog Number:
Contenido
| Item | Part # | Presentación | Concentración | Available Separately |
|---|---|---|---|---|
|
Methionine |
L118A | 1 × 50μl | 1mM | |
|
T7 TnT® PCR Enhancer |
L216A | 1 × 100μl | ||
|
Luciferase T7 Control DNA |
L482A | 1 × 5μg | 0.5μg/μl | |
|
Luciferase Assay Reagent |
L486A | 1 × 250μl | ||
|
TnT® T7 Quick Master Mix |
L520A | 8 × 200μl | ||
|
Nuclease-Free Water |
P119A | 1 × 1,250μl | View Product |
SDS
Search for SDSCertificado de Análisis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Condiciones de Almacenaje
Contenido
| Item | Part # | Presentación | Concentración |
|---|---|---|---|
|
Methionine |
L118A | 1 × 50μl | 1mM |
|
T7 TnT® PCR Enhancer |
L216A | 1 × 100μl | |
|
Luciferase T7 Control DNA |
L482A | 1 × 5μg | 0.5μg/μl |
|
TnT® T7 Quick Master Mix |
L520A | 1 × 200μl |
SDS
Search for SDSCertificado de Análisis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Condiciones de Almacenaje
Contenido
| Item | Part # | Presentación | Concentración | Available Separately |
|---|---|---|---|---|
|
Methionine |
L118A | 1 × 50μl | 1mM | |
|
TnT® SP6 Quick Master Mix |
L209A | 8 × 200μl | ||
|
Luciferase SP6 Control DNA |
L474A | 1 × 5μg | 0.5μg/μl | |
|
Luciferase Assay Reagent |
L486A | 1 × 250μl | ||
|
Nuclease-Free Water |
P119A | 1 × 1,250μl | View Product |
SDS
Search for SDSCertificado de Análisis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Condiciones de Almacenaje
Contenido
| Item | Part # | Presentación | Concentración |
|---|---|---|---|
|
Methionine |
L118A | 1 × 50μl | 1mM |
|
TnT® SP6 Quick Master Mix |
L209A | 1 × 200μl | |
|
Luciferase SP6 Control DNA |
L474A | 1 × 5μg | 0.5μg/μl |
SDS
Search for SDSCertificado de Análisis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Condiciones de Almacenaje
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