Takara Human Cell-Free Protein Expression System and vectors

Overview

• Easy-to-use system allows generation of protein in as little as 1 hour in a single-tube reaction
• Provides higher yield and greater consistency of functional protein compared to rabbit reticulocyte in vitro translation systems
• Works well with challenging applications such as large proteins (over 150 kDa)
• Amenable to high-throughput screening; bulk sizes are available

More Information

Applications

• Expression of toxic proteins that are lethal for in vivo expression systems
• Quick analysis of protein functionality
• Rapid analysis of protein mutations or truncations
• Expression of proteins that are easily degraded or insoluble in conventional in vivo expression systems such as E. coli
• High-throughput proteomic studies

Storage

• Human Cell-Free Protein Expression System (Cat. # 3281): –80°C
• pT7-IRES Vectors (Cat. # 3290, 3291, 3292): –20°C
• Human Cell-Free Protein Expression Maxi System (Cat. # 3285): Dialysis columns & accessories 4°C; All other components –80°C

Components

Human Cell-Free Protein Expression System
Cell Lysate	100 µl
Mixture-1	60 µl
Mixture-2	10 µl
Mixture-3	20 µl
T7 RNA Polymerase (200 U/µl)	10 µl
pT7 IRES Vector (0.5 µg/µl)	20 µl
Control Vector (0.3 µg/µl)	10 µl

Notes:

• Dissolve Cell Lysate just prior to use, mixing gently by pipetting. Use immediately. After use, promptly store at –80°C. Although up to five freeze-thaw cycles should not affect performance, store the Cell Lysate in small-volume, ready-to-use aliquots.
• Mixture-2 and Mixture-3 contain protein. To avoid deactivation, do not stir excessively or vortex.
• The Control Vector contains a beta-galactosidase gene cassette.
• The pT7 IRES Vector included in the kit does not encode fusion tags. Vectors encoding a His or c-myc tag are sold separately.

pT7-IRES vectors

Each vector (Cat. # 3290, 3291, 3292) contains 20 µg DNA at a concentration of 0.5 µg/µl (40 µl total volume).

Human Cell-Free Protein Expression Maxi System
1) Human Cell-Free Protein Expression System (Cat. # 3281)	2 kits
2) Supplement Buffer (Human Cell-Free Protein Expression)	15 ml
3) Dialysis Columns & Accessories	25 sets

Notes:

• Items (2) and (3) are not available for sale separately.
• Item (3) is manufactured by G-Biosciences and includes the following components:
  • Tube-O-DIALYZER™, Micro 15K MWCO (5 tubes) Note: Each tube is filled with a preservative solution. The shipping package also contains a small amount of preservative solution to maintain moisture.
  • Micro Dialysis Cup (5 cups)
  • Float for Micro, for Dialysis Cup (5 floats)

pT7-IRES vector information

Vectors in the pT7-IRES series include a T7 promoter and EMCV IRES sequence to facilitate transcription and translation in the Human Cell-Free Protein Expression System as well as a Multiple Cloning Site (MCS), convenient tags (N- or C-terminal His or N-terminal Myc tag sequences), a Factor Xa protease cleavage site for tag removal, polyA signal and T7 terminator.

• pT7-IRES His-N DNA (Cat. # 3290) encodes an N-terminal His tag
• pT7-IRES His-C DNA (Cat. # 3291) encodes a C-terminal His tag
• pT7-IRES Myc-N DNA (Cat. # 3292) encodes an N-terminal c-Myc tag

For vector maps and sequence files, see User Manuals and Vector Information, respectively, under Documents.

Related In-Fusion cloning products

For rapid and easy cloning, use the In-Fusion Cloning Systems or Kits. Availability of products in the In-Fusion portfolio varies by geographic location; see the In-Fusion product pages for more information about products available in your region.

Additional product information

Please see the product`s Certificate of Analysis and User Manual for information about storage conditions, product components, and technical specifications. Technical documents are located under the Documents tab.