Features- Easy-to-use system allows generation of protein in as little as 1 h in a single-tube reaction
- Provides higher yield of functional protein and greater consistency than rabbit reticulocyte or wheat germ in vitro translation systems
- Excellent with challenging proteins such as large proteins (over 150 kDa) and proteins requiring post-translational modification
- Amenable to high-throughput screening; bulk sizes available
Applications- Expression of toxic proteins that are lethal to host cells of in vivo expression systems
- Rapid analysis of protein function
- Rapid analysis of mutation series or truncation series: generate protein quickly and assess functionality using your downstream assay
- High-throughput proteomic studies
- Expression of proteins that are easily degraded or insoluble in conventional in vivo expression systems such as E. coli
- High-throughput proteomic studies
ComponentsHuman 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, mix gently by pipetting, and use immdiately. After use, store promptly at -80°C. Although up to five freeze-thaw cycles should not affect performance, store the Cell Lysate in aliquots of the desired volume.
- 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-tag or c-myc tag are sold separately.
pT7-IRES VectorsEach vector (Cat. #s 3290, 3291, 3292) provides 20 µg DNA at a concentration of 0.5 µg/µL (40 µL total volume).
StorageHuman Cell-Free Protein Expression System (Cat. # 3281): –80°C pT7-IRES Vectors (Cat. #s 3290, 3291, 3292): –20°C
pT7-IRES Vector informationVectors 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 tag or 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, see Data Sheets and User Manuals under "Documents". For vector sequence files, see "Resources."
Related In-Fusion Cloning productsFor rapid and easy cloning, use the In-Fusion Cloning Systems or Kits. Products in the In-Fusion portfolio vary by geographic location; see the In-Fusion product pages from Clontech for more information about products available in your region.
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