TnT® Coupled Reticulocyte Lysate Systems
A Single-Tube, Coupled Transcription/Translation System
- Study protein:protein, protein:nucleic acid and protein expression interactions
- Generate proteins in one hour versus days
- You choose the promoter
Catalog Number:
Choose a Promoter
Size
Catalog Number: L4600
Catalog Number: L4601
Catalog Number: L4610
Catalog Number: L4611
Catalog Number: L4950
Catalog Number: L5010
Catalog Number: L5020
Eukaryotic Cell-Free Protein Expression
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.
For more information about cell-free protein expression, see the Protein Expression chapter of the Protocols & Applications Guide.
Applications
- 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).
References
- Pelham, H.R.B. and Jackson, R.J. (1976) Eur. J. Biochem. 67, 247–56.
- Krieg, P.A. and Melton, D.A. (1984) Nucl. Acids Res. 12, 7057–70.
Protocols
Specifications
Catalog Number:
What's in the box
| Item | Part # | Choose a size | Concentration | Available Separately |
|---|---|---|---|---|
|
Amino Acid Mixture Minus Cysteine |
L447C | 1 × 50μl | 1mM | |
|
Luciferase Assay Wells |
L457A | 1 × 3 each | ||
|
TnT® Reaction Buffer |
L462A | 1 × 90μl | ||
|
TnT® SP6 Polymerase |
L463A | 1 × 60μl | ||
|
Luciferase SP6 Control DNA |
L474A | 1 × 5μg | 0.5μg/μl | |
|
TnT® Rabbit Reticulocyte Lysate |
L483A | 5 × 200μl | ||
|
Luciferase Assay Reagent |
L486A | 1 × 250μl | ||
|
Amino Acid Mixture Minus Leucine |
L995A | 1 × 50μl | 1mM | |
|
Amino Acid Mixture Minus Methionine |
L996A | 1 × 50μl | 1mM | View Product |
|
Magnesium Acetate |
L458A | 1 × 100μl | 25mM |
SDS
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Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
See Protocol for detailed storage recommendations.
What's in the box
| Item | Part # | Choose a size | Concentration | Available Separately |
|---|---|---|---|---|
|
Luciferase Assay Wells |
L457A | 1 × 3 each | ||
|
TnT® Reaction Buffer |
L462B | 1 × 20μl | ||
|
TnT® SP6 Polymerase |
L463C | 1 × 20μl | ||
|
Luciferase SP6 Control DNA |
L474A | 1 × 5μg | 0.5μg/μl | |
|
TnT® Rabbit Reticulocyte Lysate |
L483A | 1 × 200μl | ||
|
Luciferase Assay Reagent |
L486A | 1 × 250μl | ||
|
Amino Acid Mixture Minus Leucine |
L995D | 1 × 25μl | 1mM | |
|
Amino Acid Mixture Minus Methionine |
L996D | 1 × 25μl | 1mM | View Product |
|
Magnesium Acetate |
L458A | 1 × 100μl | 25mM |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
See Protocol for detailed storage recommendations.
What's in the box
| Item | Part # | Choose a size | Concentration | Available Separately |
|---|---|---|---|---|
|
Amino Acid Mixture Minus Cysteine |
L447C | 1 × 50μl | 1mM | |
|
Luciferase Assay Wells |
L457A | 1 × 3 each | ||
|
TnT® Reaction Buffer |
L462A | 1 × 90μl | ||
|
TnT® T7 Polymerase |
L481A | 1 × 60μl | ||
|
Luciferase T7 Control DNA |
L482A | 1 × 5μg | 0.5μg/μl | |
|
TnT® Rabbit Reticulocyte Lysate |
L483A | 5 × 200μl | ||
|
Luciferase Assay Reagent |
L486A | 1 × 250μl | ||
|
Amino Acid Mixture Minus Leucine |
L995A | 1 × 50μl | 1mM | |
|
Amino Acid Mixture Minus Methionine |
L996A | 1 × 50μl | 1mM | View Product |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
See Protocol for detailed storage recommendations.
What's in the box
| Item | Part # | Choose a size | Concentration | Available Separately |
|---|---|---|---|---|
|
Luciferase Assay Wells |
L457A | 1 × 3 each | ||
|
TnT® Reaction Buffer |
L462B | 1 × 20μl | ||
|
TnT® T7 Polymerase |
L481C | 1 × 20μl | ||
|
Luciferase T7 Control DNA |
L482A | 1 × 5μg | 0.5μg/μl | |
|
TnT® Rabbit Reticulocyte Lysate |
L483A | 1 × 200μl | ||
|
Luciferase Assay Reagent |
L486A | 1 × 250μl | ||
|
Amino Acid Mixture Minus Leucine |
L995D | 1 × 25μl | 1mM | |
|
Amino Acid Mixture Minus Methionine |
L996D | 1 × 25μl | 1mM | View Product |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
See Protocol for detailed storage recommendations.
What's in the box
| Item | Part # | Choose a size | Concentration | Available Separately |
|---|---|---|---|---|
|
Amino Acid Mixture Minus Cysteine |
L447C | 1 × 50μl | 1mM | |
|
Luciferase Assay Wells |
L457A | 1 × 3 each | ||
|
TnT® Reaction Buffer |
L462A | 1 × 90μl | ||
|
TnT® Rabbit Reticulocyte Lysate |
L483A | 5 × 200μl | ||
|
Luciferase Assay Reagent |
L486A | 1 × 250μl | ||
|
TnT® T3 Polymerase |
L493A | 1 × 60μl | ||
|
Luciferase T3 Control DNA |
L494A | 1 × 5μg | 0.5μg/μl | |
|
Amino Acid Mixture Minus Leucine |
L995A | 1 × 50μl | 1mM | |
|
Amino Acid Mixture Minus Methionine |
L996A | 1 × 50μl | 1mM | View Product |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
See Protocol for detailed storage recommendations.
What's in the box
| Item | Part # | Choose a size | Concentration | Available Separately |
|---|---|---|---|---|
|
Amino Acid Mixture Minus Cysteine |
L447C | 1 × 50μl | 1mM | |
|
Luciferase Assay Wells |
L457A | 1 × 3 each | ||
|
TnT® Reaction Buffer |
L462A | 1 × 90μl | ||
|
TnT® T7 Polymerase |
L481B | 1 × 30μl | ||
|
Luciferase T7 Control DNA |
L482A | 1 × 5μg | 0.5μg/μl | |
|
TnT® Rabbit Reticulocyte Lysate |
L483A | 5 × 200μl | ||
|
Luciferase Assay Reagent |
L486A | 1 × 250μl | ||
|
TnT® T3 Polymerase |
L493B | 1 × 30μl | ||
|
Luciferase T3 Control DNA |
L494A | 1 × 5μg | 0.5μg/μl | |
|
Amino Acid Mixture Minus Leucine |
L995A | 1 × 50μl | 1mM | |
|
Amino Acid Mixture Minus Methionine |
L996A | 1 × 50μl | 1mM | View Product |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
See Protocol for detailed storage recommendations.
What's in the box
| Item | Part # | Choose a size | Concentration | Available Separately |
|---|---|---|---|---|
|
Amino Acid Mixture Minus Cysteine |
L447C | 1 × 50μl | 1mM | |
|
Luciferase Assay Wells |
L457A | 1 × 3 each | ||
|
TnT® Reaction Buffer |
L462A | 1 × 90μl | ||
|
TnT® SP6 Polymerase |
L463B | 1 × 30μl | ||
|
Luciferase SP6 Control DNA |
L474A | 1 × 5μg | 0.5μg/μl | |
|
TnT® T7 Polymerase |
L481B | 1 × 30μl | ||
|
Luciferase T7 Control DNA |
L482A | 1 × 5μg | 0.5μg/μl | |
|
TnT® Rabbit Reticulocyte Lysate |
L483A | 5 × 200μl | ||
|
Luciferase Assay Reagent |
L486A | 1 × 250μl | ||
|
Amino Acid Mixture Minus Leucine |
L995A | 1 × 50μl | 1mM | |
|
Amino Acid Mixture Minus Methionine |
L996A | 1 × 50μl | 1mM | View Product |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
See Protocol for detailed storage recommendations.
Resources
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