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Rabbit Reticulocyte Lysate Cell-Free Protein Expression

Rabbit Reticulocyte Lysate Cell-Free Protein Expression

Cell-free protein synthesis (CFPS) systems have been widely used for decades as a fast and effective tool in basic biology. Profacgen has developed a variety of protein expression systems using cell extracts, such as E. coli cell-free expression system, wheat germ cell-free expression system, rabbit reticulocyte lysate cell-free expression system, and tobacco cell-free expression system. Among them, Rabbit reticulocyte lysate, which reticulocytes are highly specialized cells primarily responsible for the synthesis of hemoglobin, is an efficient in vitro eukaryotic protein synthesis system used to translate foreign RNA (naturally or in vitro generated RNA).

The RNA used in the standard rabbit reticulocyte lysate translation system is derived from transcription products synthesized in vitro using SP6, T3, and T7 RNA polymerase promoters, requiring three separate reactions, and requiring multiple steps between each reaction.

Protocal of the rabbit reticulocyte lysate cell-free protein expression system:

  1. Preparation of Rabbit reticulocyte lysate;
  2. Add amino acid, SP6, T3 or T7 RNA polymerase and energy substance to form the reaction system;
  3. Add plasmid DNA or PCR products, synthesize mRNA in vitro under the action of RNA polymerase, incubate and detect;
  4. Use transcription factors in cell extracts, enzymes required for various synthetic proteins, and supplementary amino acids, energy substances, tRNA, etc. to translate mRNA into proteins and purify the target protein.

Profacgen can now provide you with a TNT-coupled reticulocyte lysate system with shorter translation time and higher protein expression than traditional rabbit reticulocyte lysate translation systems. This is a single-tube, coupled transcription / translation system, which refers to the incorporation of transcription products directly into the translation mixture (including RNA polymerase, nucleotides, salts, RNasin nuclease inhibitors), only need to add templates (PCR fragments or plasmids) and methionine (generally radioisotope labels) to complete the entire translation process. At the same time we can provide different concentrations of Mg2+ and K+ and the option of adding DTT to optimize the translation process.

Advantages of the rabbit reticulocyte lysate cell-free protein expression system:

  1. Extract preparation system is simple and cost-effective;
  2. Easy cell breakage and quick preparation of extract;
  3. High protein yield of hemoglobin protein while moderate/low yields for other proteins;
  4. High protein synthesis rate, which can achieve the simultaneous expression of multiple proteins in the same system

Profacgen's cell-free expression platform can provide you with comprehensive technical services, specializing in the special properties of the target protein, and professionally solve protein expression related problems, such as low protein production, expression of special proteins (such as membrane proteins, toxic proteins, etc.), and preparation of proteins Complex, parallel synthesis of many different proteins, etc. We always put your needs first, and if you have any questions about the results, we can help you verify. Profacgen is your ideal partner. If you are interested in our services, please feel free to contact us for more information

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