|
New products
|
Cell-Free Expression with AvidExpressTM Translation System
|
AvidExpress™ cell-free translation system 
Translation initiation in mammalian cells is a complex process that requires a set of specific eukaryotic initiation factors for positioning 40S ribosomal subunit at the specific translation site. Failure of this process results in aberrant initiation, premature termination or miscoding during protein synthesis. The newly synthesized proteins also require eukaryotic chaperons and membranous structure for correct folding. Contrary to bacterial and plant derived cell-free lysates, our AvidExpress™ cell-free translation system provides human HeLa S3 cell-derived translation factors, amino acids, chaperons and membranes for translation of cellular or in vitro synthesized mRNAs.
Features:
- Human cell lysate-based translation system
- The method is simple: just add supplied components and your RNA template.
- Just as with natural mRNAs, in vitro transcribed RNAs containing internal ribosome entry site (IRES) or cap structure at the 5’ end and poly(A) tail at the 3’end can be efficiently translated.
- The translation products can be easily labeled with [35S]Methionine or biotinylated amino acids or use our AviTag sequence in your mRNA for detection of translated protein.
- Lysates can be used to study protein-protein, RNA-protein and DNA-protein interactions by UV cross-linking, electrophoresis mobility shift assay (EMSA) and immunoprecipitation assays.
- The system can be scaled-up easily.
-
Biotinylated proteins can be synthesized for direct immobilization on solid surfaces coated with streptavidin or its molecular analogues |
|
Kinetics of an IRES-contrlolled translation in AvidExpress™ cell-free translation system (graph) of firefly luciferase. |
 SDS-PAGE of translated protein followed by detection of protein band with chemiluminescence method. Lane 1, control lysate without exogenous RNA; lane 2, in vitro transcribed RNA expressing firefly luciferase. |
Reference:
1 Barton, D. J., B. J. Morasco, and J. B. Flanegan. 1996. Assays for poliovirus polymerase, 3D(Pol), and authentic RNA replication in HeLa S10 extracts. Methods Enzymol. 275:35-57. |
|
|