Thermo Scientific 1-Step Heavy Protein in vitro Protein Expression Kits enable rapid cell-free expression of recombinant proteins containing stable isotope-labeled (i.e., heavy) amino acids.
Features of the 1-Step Heavy Protein IVT Kit:
• Efficient—express heavy proteins with 90 to 95% stable isotope incorporation • Functional—uses the human translational machinery to express more biologically active proteins than other IVT systems • Flexible—express light proteins or heavy proteins containing 13C615N2 L-lysine and/or 13C615N4 L-arginine • Convenient—perform transcription and translation in a single step • High performance—greater yields compared to rabbit reticulocyte in vitro translation
The 1-Step Heavy Protein IVT Kit uses a unique HeLa cell lysate supplemented with heavy amino acids for in vitro translation (IVT) of proteins with 90 to 95% isotope incorporation efficiency in less than 8 hours. Heavy proteins expressed using this system can be used as mass spectrometry controls for sample prep loss, digestion efficiency determination or as quantification standards.
Applications: • Identify ideal peptides for targeted quantitation • Verify protein digestion efficiency • Control for protein sample prep variability and affinity enrichment loss
Thermo Scientific 1-Step Heavy Protein IVT Kit contains all of the necessary components to express a heavy protein including HeLa cell lysate, proprietary accessory proteins, reaction mix, heavy amino acids, positive-control GFP DNA and the pT7CFE1-CGST-HA-His cloning vector. The benefits of in vitro expression of heavy proteins over traditional in vivo systems include expression of toxic or insoluble proteins, a more rapid protein synthesis and a more economical use of heavy amino acids compared to stable-isotope labeled cell lines. This small-scale expression method makes it easy to express numerous heavy proteins simultaneously or to express large quantities of a single heavy protein up to 100 µg/mL. Tandem affinity purification of heavy proteins is aided using an expression vector containing multiple affinity tags including GST, HA and 6xHis.