General description

Cell-free protein expression (CFPE) systems derived from crude cell extracts have been used for decades as a research tool and have introduced many attractive advantages to protein expression technology. However, a critical barrier to the adoption of CFPE systems as alternatives to cell-based approaches has been the problem of relatively low protein yields.

To address this problem, ALiCE^??a novel eukaryotic cell-free expression system was created by taking advantage of plant cell biology. ALiCE^??offers unprecedented yields of up to 3 mg/mL of protein in batch mode. As such, it enables you to overcome the limitations of living host systems and to obtain “difficult to produce” proteins in a matter of hours instead of weeks.

ALiCE^??utilizes tobacco cell lysates, depleted of vacuoles. The system contains all factors required for in vitro transcription (RNA polymerases, NTPs) and translation reactions (ribosomes, translation initiation/elongation factors, tRNA, etc.). Using a simple protocol, both the RNA transcription and translation take place in a single tube (coupled transcription/translation).

Reactions are started by simply mixing plasmid DNA with the ALiCE^??Reaction mix. The outstanding protein yields of ALiCE^??are based on highly efficient protein expression over the duration of the reaction. Consequently, we recommend a reaction duration of 48 h.

ALiCE^??is provided with two expression vectors, pALiCE01 and pALiCE02, which are used according to the targeting of the protein of interest. These have been specially tailored to offer excellent protein yield when used with the ALiCE^??expression system. pALiCE01 is used for expression in ALiCE^??cytosolic fraction. Targeting the microsomes with pALiCE02 offers the opportunity to express protein with post-translational modifications like disulfide bonds.

Application

Besides screening the ALiCE^??system can also be used for larger scale expression of target proteins. Potential additional applications of the ALiCE^??system include:

??Target protein characterization
??Protein optimization
??Mutant screening
??Protein-protein interactions
??Expression analysis
??Protein localization analysis
??Protein structure analysis
??Post-translational modification assays
??Metabolomics
??Herbicide screening

Features and Benefits

ALiCE^??Cell Free Protein Expression Kit?utilizes cell lysate derived from plant cells to produce recombinant protein at 3 mg/mL in batch mode.
??High-yield: Up to 3 mg/mL expressed protein
??Easy to use: One-step reaction protocol
??Fast: High yield in just 48 hours
??Flexible: Express “difficult to produce” proteins, cytotoxic proteins, etc.

Components

ALiCE^??Mini Kit?-?AL0103050?Comprises
ALiCE^??Lysate, 6 x 50 μL
Vector pALiCE01, 15 μL
Vector pALiCE02, 15 μL
ALiCE^??Tube Cap, 0.6 mm hole

ALiCE^??Midi Kit?-?AL0103200?Comprises
ALiCE^??Lysate, 6 x 200 μL
Vector pALiCE01, 50 μL
Vector pALiCE02, 50 μL
ALiCE^??Tube Set, 12 pcs

ALiCE^??Maxi Kit?-?AL0103500?Comprises
ALiCE^??Lysate, 6 x 500 μL
Vector pALiCE01, 50 μL
Vector pALiCE02, 50 μL
ALiCE^??Tube Set, 12 pcs

Storage and Stability

??ALiCE^??Reaction (Lysate) mix: store at -70 °C
??Vector pALiCE01: store at -20 °C to -70 °C
??Vector pALiCE02: store at -20 °C to -70 °C
??ALiCE^??Tube caps/Tube set: store at room temperature

法律信息

ALiCE is a registered trademark of LenioBio GmbH