Cell Signaling TNT® Quick Coupled Transcription/Translation System Product TN TN Size Conc. Cat.# Price (Fr) T® T7 Quick Coupled Transcription/ Translation System T® T7 Quick Coupled Transcription/ Translation System, Trial Size TNT® SP6 Quick Coupled Transcription/Translation System TNT® SP6 Quick Coupled Transcription/Translation System, Trial Size Magnesium Acetate Potassium Chloride 40 reactions 5 reactions L2080 1016.00 L2081 254.00 100 µl 25 mM L4581 85.00 200 µl 2.5 M L4591 85.00 L4581, L4591 For Laboratory Use. L1170, L1171, L2080, L2081 For Research Use Only. Not for Use in Diagnostic Procedures. Description: 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 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. Features: • Obtain Data Faster: Functional protein is expressed in only one hour, not days as with cell-based expression systems. • Multiple Applications with One System: Use expressed protein for the characterization of protein:protein interaction, protein:nucleic acid interaction, protein modification and more. • Consistent, Reliable Results: This mammalian-based system expresses soluble, functional proteins that are post-translationally modified, unlike E. coli-based systems. • Fewer Steps: Expressed proteins can be used directly after expression; no requirement for additional purification. • Flexible Systems Available: TNT® Systems for linear, circular or PCR templates are available. Storage Conditions: Store at –70°C. Do not freeze-thaw the lysate more than two times. Protocol TNT® Quick Coupled Transcription/Translation System Technical Manual Part# TM045 40 reactions 5 reactions L1170 1016.00 L1171 254.00 TNT® Coupled Reticulocyte Lysate Systems Product T® SP6 Coupled Reticulocyte Lysate System, Trial Size Size Cat.# Price (Fr) TNT® SP6 Coupled Reticulocyte Lysate System 40 reactions L4600 996.00 TN 8 reactions L4601 249.00 T® T7 Coupled Reticulocyte Lysate System, Trial Size T® T7/T3 Coupled Reticulocyte Lysate System TN TNT® T7 Coupled Reticulocyte Lysate System 40 reactions L4610 996.00 TN 8 reactions L4611 249.00 TNT® T3 Coupled Reticulocyte Lysate System 40 reactions L4950 996.00 TN 40 reactions L5010 1046.00 T® T7/SP6 Coupled Reticulocyte Lysate System 40 reactions L5020 1046.00 For Research Use Only. Not for Use in Diagnostic Procedures. Description: The TNT® Coupled Reticulocyte Lysate Systems offer researchers an alternative for eukaryotic cell-free protein expression: a singletube, 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. Features: • 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. Storage Conditions: Store the polymerase at –20 to –70°C. Store Luciferase Assay Wells at room temperature. Store the other components at –70°C. Do not freeze-thaw the lysate more than two times. Protocol TNT® Coupled Reticulocyte Lysate System Technical Bulletin Part# TB126 13 For complete and up-to-date product information visit: www.promega.com/catalog 225 Protein Expression and Analysis Pagina 228
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