Shop All Protein Expression Kits

Pichia Expression Kit, original kit (Invitrogen™)

The Original Pichia Expression Kit is designed for high-level expression of recombinant protein in the yeast Pichia pastoris. The vectors carry the HIS4 gene for selection of transformants on histidine-deficient medium. The Original Pichia Expression Kit provides all of the tools and reagents needed to express a gene of interest from the AOX1 promoter.

FreeStyle™ 293 Expression System (Invitrogen™)

The FreeStyle™ 293 Expression System is a complete, suspension cell culture system for generating large amounts of mammalian recombinant protein. Free your lab from the time-consuming, labor-intensive process of seeding, feeding, passing, and maintaining multiple flasks of adherent cells (Figure 1). The FreeStyle™ 293 Expression System combines GIBCO® FreeStyle™ 293 Expression Medium with 293fectin™, a cationic-lipid formulation designed to transfect suspension FreeStyle™ 293-F cells at high efficiency. FreeStyle™ 293-F cells and 293fectin™ are available as part of the Expression System or separately.

FreeStyle™ 293 Expression Medium is chemically-defined, protein-free medium specifically developed for the ability to support the growth and transfection of 293-F cells under suspension type culture conditions (Figure 2). The medium is a complete, ready-to-use medium that has been supplemented with GlutaMAX™-I Supplement and is animal-origin free. FreeStyle™ 293 Expression Medium is able to save significant time and costs associated with adaptation of cell cultures to serum-free conditions.

pBAD TOPO™ TA Expression Kit (Invitrogen™)

The pBAD TOPO® TA Expression Kit is specifically designed for one-step cloning and regulated prokaryotic expression of Taq-amplified PCR products. Once you've TOPO® Cloned your PCR product, you can go straight to protein expression. Some of the convenient features of the pBAD-TOPO® vector include:

• Linearized, topoisomerase I-activated vector for 5-minute cloning of Taqamplified PCR products
• The araBAD promoter for tightly regulated expression in E. coli V5 epitope tag for detection with an Anti-V5 Antibody
• C-terminal polyhistidine (6xHis) tag for purification using nickel-chelating resin and detection with an Anti-His(C-term) Antibody
• Enterokinase cleavage site for removal of N-terminal leader peptide

FreeStyle™ MAX 293 Expression System (Gibco™)

The FreeStyle™ MAX System is a breakthrough technology for rapid and high-yield mammalian protein production. This new mammalian expression technology is available in two convenient formats. FreeStyle™ MAX Reagent is a novel cationic lipid-based reagent designed to transfect FreeStyle™ cells with high efficiency. The FreeStyle™ MAX Expression System combines GIBCO® FreeStyle™ Media, FreeStyle™ MAX Reagent, and either FreeStyle™ CHO-S cells or FreeStyle™ 293 cells. The only equipment required to use the FreeStyle™ MAX System is a CO2 incubator and a platform shaker with shake flasks or a spinner culture.

The FreeStyle™ MAX System provides:
• An easy, rapid protocol for transient transfection of CHO and 293 cells in suspension cultures
• Functional proteins (up to milligram quantities) with all post-translational modifications produced in one week
• Simplified downstream purification of secreted proteins with FreeStyle™ serum-free medium
• Straightforward protocol for production scale-up

The FreeStyle™ MAX System can replace time-consuming stable cell line generation with large-scale functional protein production in one week.

Contents and Storage:
Store FreeStyle™ cells in liquid nitrogen. Store FreeStyle™ MAX Reagent at +4°C. Store GIBCO® FreeStyle™ CHO Expression Medium, FreeStyle™ 293 Expression Medium, and OptiPRO™ SFM at +2-8°C in the dark. Store the pCMV SPORT at -20°C. Stability is guaranteed for 6 months under the recommended storage conditions. Please refer to the manual for additonal storage information.

Bac-to-Bac™ C-His TOPO™ Expression System (Gibco™)

Bac-to-Bac baculovirus expression is an efficient method for producing baculovirus for high-level protein expression in insect cells. It relies on generation of recombinant virus by site-specific transposition in E. coli rather than homologous recombination in insect cells. The pFastBac C-His TOPO expression system features:

Flexibility—The Bac-to-Bac C-His TOPO vector contains a C-terminal His-tag with a TEV cleavage site for easy purification of His- tagged proteins with nickel-chelating resins (such as the Invitrogen ProBond Purification System) and generation of native proteins with the aid of the Invitrogen AcTEV Protease.

Fast cloning, easy screening—Blunt TOPO cloning technology enables cloning of blunt-end PCR products into the pFastBac TOPO vector in five minutes. In addition, the pFastBac TOPO expression kit is supplied with Mach1-T1R E. coli that enable the visualization of colonies eight hours after plating on ampicillin-selective plates due to their faster doubling time compared to other standard cloning strains. Find out more about the advantages of TOPO cloning ›

High transfection efficiency with ExpiFectamine Sf Transfection Reagent—The Bac-to-Bac TOPO expression kits now come with the next-generation ExpiFectamine Sf Transfection Reagent for efficient DNA transfection in insect cells using fast, flexible protocols. Find out more about this reagent ›

Time-saving expression bacmid—With the Bac-to-Bac system, the expression cassette of the pFastBac vector recombines with the parent bacmid in DH10Bac E. coli Competent Cells to form an expression bacmid. The bacmid is then transfected into insect cells for production of recombinant baculovirus particles.

Easy colony screening—The parent bacmid in DH10Bac E. coli contains a segment of the lacZa gene. The lacZa gene is disrupted upon transposition of the expression cassette into the bacmid, allowing for blue/white selection of recombinants for easier identification of recombinant colonies.

High protein expression—The pFastBac vector uses the strong polyhedrin promoter to generate high levels of expression in a variety of insect cell line such as Sf9, Sf21, and High Five cells.

pTrcHis2 TOPO™ TA Expression Kit (Invitrogen™)

The pTrcHis- and pTrcHis2-TOPO® TA Expression Kits allow fast, efficient cloning of PCR products into a prokaryotic expression vector for high-level, regulated expression from the trc promoter. The vector in each kit, pTrcHis-TOPO® or pTrcHis2-TOPO®, includes a minicistron element for enhanced translation efficiency of eukaryotic proteins in E. coli. Both vectors are provided linearized and activated with topoisomerase I for 5-minute TOPO® Cloning with >85% cloning efficiency. To simplify protein purification and detection, the pTrcHis-TOPO® and pTrcHis2-TOPO® vectors include the following features:

pTrcHis-TOPO®

N-terminal Xpress™ epitope for detection with an Anti-Xpress™ Antibody
• N-terminal polyhistidine (6xHis) tag for purification using nickelchelating resin and detection with an Anti-HisG Antibody
• Enterokinase cleavage site for efficient removal of N-terminal fusion tags

pTrcHis2-TOPO®
C-terminal c-myc epitope for detection with an Anti-myc Antibody
• C-terminal polyhistidine (6xHis) tag for purification using nickel-chelating resin and detection with an Anti-His(C-term) Antibody

FreeStyle™ MAX CHO Expression System (Gibco™)

The FreeStyle™ MAX System is a breakthrough technology for rapid and high-yield mammalian protein production. This new mammalian expression technology is available in two convenient formats. FreeStyle™ MAX Reagent is a novel cationic lipid-based reagent designed to transfect FreeStyle™ cells with high efficiency. The FreeStyle™ MAX Expression System combines GIBCO® FreeStyle™ Media, FreeStyle™ MAX Reagent, and either FreeStyle™ CHO-S cells or FreeStyle™ 293 cells. The only equipment required to use the FreeStyle™ MAX System is a CO2 incubator and a platform shaker with shake flasks or a spinner culture.

The FreeStyle™ MAX System provides:
• An easy, rapid protocol for transient transfection of CHO and 293 cells in suspension cultures (Figure 1)
• Functional proteins (up to milligram quantities) with all post-translational modifications produced in one week
• Simplified downstream purification of secreted proteins with FreeStyle™ serum-free medium
• Straightforward protocol for production scale-up

The FreeStyle™ MAX System can replace time-consuming stable cell line generation with large-scale functional protein production in one
week (Figure 2).

Contents and Storage:
Store FreeStyle™ cells in liquid nitrogen. Store FreeStyle™ MAX Reagent at +4°C. Store GIBCO® FreeStyle™ CHO Expression Medium, FreeStyle™ 293 Expression Medium, and OptiPRO™ SFM at +2-8°C in the dark. Store the pCMV SPORT at -20°C. Stability is guaranteed for 6 months under the recommended storage conditions. Please refer to the manual for additonal storage information.

Bac-to-Bac™ N-His TOPO™ Expression System (Gibco™)

Bac-to-Bac baculovirus expression is an efficient method for producing baculovirus for high-level protein expression in insect cells. It relies on generation of recombinant virus by site-specific transposition in E. coli rather than homologous recombination in insect cells. The pFastBac N-His TOPO expression system features:

Flexibility—The Bac-to-Bac N-His TOPO vector contains an N-terminal His-tag with a TEV cleavage site to enable easy purification of His-tagged proteins with nickel-chelating resins (such as the Invitrogen ProBond Purification System) and generation of native proteins with the aid of the Invitrogen AcTEV Protease.

Fast cloning, easy screening—Blunt TOPO cloning technology enables cloning of blunt-end PCR products into the pFastBac TOPO vector in five minutes. In addition, the pFastBac TOPO expression kit is supplied with Mach1-T1R E. coli that enable the visualization of colonies eight hours after plating on ampicillin-selective plates due to their faster doubling time compared to other standard cloning strains. Find out more about the advantages of TOPO cloning ›

High transfection efficiency with ExpiFectamine Sf Transfection Reagent—The Bac-to-Bac TOPO expression kits now come with the next-generation ExpiFectamine Sf Transfection Reagent for efficient DNA transfection in insect cells using fast, flexible protocols. Find out more about this reagent ›

Time-saving expression bacmid—With the Bac-to-Bac system, the expression cassette of the pFastBac vector recombines with the parent bacmid in DH10Bac E. coli Competent Cells to form an expression bacmid. The bacmid is then transfected into insect cells for production of recombinant baculovirus particles.

Easy colony screening—The parent bacmid in DH10Bac E. coli contains a segment of the lacZa gene. The lacZa gene is disrupted upon transposition of the expression cassette into the bacmid, allowing for blue/white selection of recombinants for easier identification of recombinant colonies.

High protein expression—The pFastBac vector uses the strong polyhedrin promoter to generate high levels of expression in a variety of insect cell line such as Sf9, Sf21, and High Five cells.

pTrcHis TOPO™ TA Expression Kit (Invitrogen™)

The pTrcHis- and pTrcHis2-TOPO® TA Expression Kits allow fast, efficient cloning of PCR products into a prokaryotic expression vector for high-level, regulated expression from the trc promoter. The vector in each kit, pTrcHis-TOPO® or pTrcHis2-TOPO®, includes a minicistron element for enhanced translation efficiency of eukaryotic proteins in E. coli. Both vectors are provided linearized and activated with topoisomerase I for 5-minute TOPO® Cloning with >85% cloning efficiency. To simplify protein purification and detection, the pTrcHis-TOPO® and pTrcHis2-TOPO® vectors include the following features:

pTrcHis-TOPO®

N-terminal Xpress™ epitope for detection with an Anti-Xpress™ Antibody
• N-terminal polyhistidine (6xHis) tag for purification using nickelchelating resin and detection with an Anti-HisG Antibody
• Enterokinase cleavage site for efficient removal of N-terminal fusion tags

pTrcHis2-TOPO®
C-terminal c-myc epitope for detection with an Anti-myc Antibody
• C-terminal polyhistidine (6xHis) tag for purification using nickel-chelating resin and detection with an Anti-His(C-term) Antibody

Champion™ pET160 Directional TOPO™ Expression Kit with Lumio™ Technology (Invitrogen™)

The Champion™ pET Expression System yields the highest-level protein production in E. coli. During expression, your protein of interest can reach levels greater than 50 percent of total cellular protein. Based on T7 expression vectors originally developed by Studier and colleagues (1-3), high-level expression is achieved because the T7 RNA polymerase is more processive than native E. coliRNA polymerase and is dedicated to the transcription of your gene of interest. Protein production is further enhanced in the system by the expression strain BL21 Star™ E. coli, which significantly improves the stability of mRNA transcripts and increases protein expression up to ten-fold.

Simplified, efficient Directional TOPO® cloning allows you to quickly enter a Champion™ pET Expression vector. These kits feature linearized, topoisomerase I-activated Champion™ pET expression vectors for five-minute directional cloning. Directional TOPO® Cloning technology facilitates gene expression because:

• A proofreading enzyme is used for PCR, resulting in fewer errors in cloned genes
• Greater than 90% of the clones are in the correct orientation for gene expression, reducing the time spent on colony screening

Seven Champion™ pET Directional TOPO® Expression Vectors are available (Figure 1 and Table 1): Each vector carries a T7lac promoter for high-level expression. Flexible options for simplifying protein detection, cleaving purification tags, selecting plasmid carrying clones, and/or improving protein yields are available.
With the Champion™ pET Directional TOPO® vectors you can expect highest-level protein production. Figure 2 shows expression of the lacZ gene in Champion™ pET Directional TOPO® vectors. Figure 3 demonstrates efficient cleavage using TEV protease of the N-terminal tag of a β-galactosidase fusion protein expressed from pET151/D-TOPO®.

Champion™ pET151 Directional TOPO™ Expression Kit with BL21 Star™ (DE3) One Shot™ Chemically Competent E. coli (Invitrogen™)

The Champion™ pET Expression System yields the highest-level protein production in E. coli. During expression, your protein of interest can reach levels greater than 50 percent of total cellular protein. Based on T7 expression vectors originally developed by Studier and colleagues (1-3), high-level expression is achieved because the T7 RNA polymerase is more processive than native E. coliRNA polymerase and is dedicated to the transcription of your gene of interest. Protein production is further enhanced in the system by the expression strain BL21 Star™ E. coli, which significantly improves the stability of mRNA transcripts and increases protein expression up to ten-fold.

Simplified, efficient Directional TOPO® cloning allows you to quickly enter a Champion™ pET Expression vector. These kits feature linearized, topoisomerase I-activated Champion™ pET expression vectors for five-minute directional cloning. Directional TOPO® Cloning technology facilitates gene expression because:

• A proofreading enzyme is used for PCR, resulting in fewer errors in cloned genes
• Greater than 90% of the clones are in the correct orientation for gene expression, reducing the time spent on colony screening

Seven Champion™ pET Directional TOPO® Expression Vectors are available (Figure 1 and Table 1): Each vector carries a T7lac promoter for high-level expression. Flexible options for simplifying protein detection, cleaving purification tags, selecting plasmid carrying clones, and/or improving protein yields are available.
With the Champion™ pET Directional TOPO® vectors you can expect highest-level protein production. Figure 2 shows expression of the lacZ gene in Champion™ pET Directional TOPO® vectors. Figure 3 demonstrates efficient cleavage using TEV protease of the N-terminal tag of a β-galactosidase fusion protein expressed from pET151/D-TOPO®.

Multi-Copy Pichia Expression Kit (Invitrogen™)

The Multi-Copy Pichia Expression Kit contains the pPIC3.5K, pPIC9K, and pAO815 vectors for production and selection of Pichia strains that contain more than one copy of the gene of interest. In many cases, increased copy number has led to increased expression levels.

pPIC9K and pPIC3.5K
The pPIC9K and pPIC3.5K vectors carry the kanamycin resistance gene that confers resistance to Geneticin® Reagent in Pichia. Spontaneous generation of multiple insertion events can be identified by resistance to increased levels of Geneticin® Reagent. Pichia transformants are selected on histidine-deficient medium and screened for their level of resistance to Geneticin® Reagent. The ability to grow in high concentrations of Geneticin® indicates that multiple copies of the kanamycin resistance gene and the gene of interest are integrated into the genome (1). The pPIC9K vector directs secretion of expressed proteins while proteins expressed from pPIC3.5K remain intracellular.

pAO815
pAO815 is a Pichia expression vector designed to clone multiple copies of a gene into a single vector. The vector contains a Bgl II site upstream of the 5´ AOX1 gene and a unique BamH I site downstream of the 3´ AOX1 transcription termination (TT) signal. Four steps are required to generate multiple copies of the gene of interest:
1. The gene is cloned into the unique EcoR I site in the vector.
2. The construct is digested with BamH I and Bgl II to release the "expression cassette" containing the AOX1 promoter, gene of interest, and 3´ AOX1 TT.
3. Concatemers of the expression cassette are generated by ligation in vitro.
4. The multiple copies are inserted back into the pAO815 vector and transformed into Pichia.

These vectors are also available separately.

ExpiCHO™ Expression System Kit (Gibco™)

The ExpiCHO™ Expression System Kit combines the power of rapid, ultra–high-yield transient protein production in suspension culture with the benefit of expression in Chinese hamster ovary (CHO) cells. It is based on high-density culture of ExpiCHO-S™ cells in ExpiCHO™ Expression Medium. Transient expression is powered by ExpiFectamine™ CHO transfection reagent, designed specifically for high-density CHO suspension culture, in combination with a specialized transfection enhancer and culture feed. All components work together synergistically to generate 2- to 10-fold higher protein yields than the HEK293-based Expi293™ Expression System Kit and 20- to 150-fold higher yields than the FreeStyle™ MAX CHO Expression System. Expression yields of 1 to 3 g per L of transfected culture have been demonstrated for some antibody and non-antibody proteins.

The ExpiCHO Expression System offers:

• Milligram to multigram yields of recombinant protein per liter of culture in 5 to 14 days post-transfection, which are frequently higher than transient 293-based systems
• Cost-effective transient expression in CHO cells, allowing scientists developing protein biotherapeutics to work with CHO-expressed proteins from start to finish in the drug development process
• A robust alternative mammalian transient expression host, allowing for production of proteins that are difficult to express in HEK293 cells
• Flexible, reliable culture and transfection protocols with multiple options that readily fit into current transient expression workflows
• Scalable expression for cultures from <1 mL to >10 L, thus generating the amount of protein needed for your application

The components included in the ExpiCHO Expression System Kit are: 2 vials of frozen ExpiCHO-S cells, 1 L of ExpiCHO Expression Medium, 1 ExpiFectamine CHO Transfection Kit sufficient to transfect 1 L of culture, OptiPRO™ SFM, and an antibody-expressing positive control vector. All components of the ExpiCHO Expression System Kit are also available for purchase separately. All components of the system are animal origin–free.

1-Step Human High-Yield Maxi IVT Kit (Thermo Scientific™)

The Thermo Scientific 1-Step Human High-Yield In Vitro Translation (IVT) Kits enable the expression of functional proteins with 10 to 100 times greater yield per mL than other mammalian IVT systems.

The 1-Step High-Yield IVT System uses modified HeLa cell extracts to take advantage of the robust human translation machinery and generate functional full-length proteins. In this system, protein expression is performed in a proprietary dialysis device that allows a continuous supply of nucleotides, amino acids and energy generating substrates into the reaction while removing inhibitors of proteins synthesis. This continuous-exchange cell-free (CECF) system enables protein expression in an overnight incubation of up to 750 µg/mL. The complete maxi-scale kit includes all the components required for transcription and translation of a recombinant gene, including an optimized expression vector.

Features of the 1-Step Human High-Yield Maxi IVT Kit:

High expression—up to 750 µg/mL of expressed protein
Reproducibility—low variability between experiments
Fast—express high levels of protein with 6 hours to overnight incubation
Functional—obtain functionally active proteins, including those containing disulfide bonds

1-Step Protein Expression:
Easy-to-use—transcription and translation is performed in one reaction step
Adapt for labeling—amenable to incorporation of heavy or unnatural amino acids

Includes:
• Complete kits contain cell lysate, reaction mix, accessory proteins, 5X dialysis buffer, control DNA, cloning vector, dialysis devices and centrifuge tubes

Requires:
• 30°C incubator or water bath (shaker incubator increases expression by 30 to 50%)

Applications:
• Production of recombinant proteins for structural analysis
• Generate active proteins for protein interaction studies
• Perform rapid expression and characterization of mutant proteins
• Small-scale production of viral particles
• Incorporation of unnatural amino acids or toxic protein production

The 1-Step Human High-Yield IVT Kits are cell-free protein expression systems that provide all of the essential components required for transcription and translation. The kits are optimized with Accessory Proteins and Reaction Mixes that support protein synthesis using a DNA template. The advantages of using the 1-Step Human High-Yield IVT Kits over traditional in vivo expression systems include the ability to express toxic or insoluble proteins, easily perform protein labeling with modified amino-acids and reduce the time and cost of expressing human proteins in tissue culture cells. 1-Step Human IVT Kits are optimized for use with the pT7CFE1 expression vector that utilizes the T7 viral promoter and an EMCV Internal Ribosome Entry Site (IRES) to facilitate high levels of in vitro protein expression. Using a pT7CFE vector is critical for obtaining high expression levels. For added convenience, at family of pT7CFE vectors are available with tandem affinity tags for facilitate protein detection and purification. (For smaller scale and screening options, consider the 1-Step Human Coupled IVT Kit.)

More Product Data
Choosing a vector and purification method for in vitro protein expression
System for high-throughput, high-yield in vitro translation
Expression of highly active proteins using a cell-free human IVT system

1-Step Human Coupled IVT Kit - DNA (Thermo Scientific™)

Thermo Scientific 1-Step Human In Vitro Protein Expression Kits enable the translation and post-transcriptional modification of full-length proteins from mRNA or plasmid templates with yields of up to 100 µg/mL per reaction.

The Human IVT Kits are unique HeLa cell lysate-based protein expression systems for in vitro translation or coupled transcription/translation reactions. Protein expression is performed in a single 90-minute reaction that can be extended for up to 6 hours with continued protein production up to 100 µg/mL when combined with the optimized pT7CFE1 Expression Vector. The Human IVT Kits can express functional enzymes, phosphoproteins, glycoproteins and membrane proteins for immediate use in studying protein interactions, performing rapid mutational analysis and measuring activity.

Features of the 1-Step Human Coupled IVT Kit:

Functional—uses the human translational machinery to express active proteins
Convenient—perform transcription and translation in a single step
High performance—greater yields compared to rabbit reticulocyte in vitro translation
Reliable—express proteins that fail in rabbit reticulocyte systems

Better Than Traditional Methods:
• HeLa cell-free extracts are capable of expressing proteins with post-translational modifications
• Accurate translation delivers full-length protein compatible with downstream applications
• Protein translation is optimized with EMCV IRES element and other mRNA stabilizing elements

Includes:
• Complete kits contain HeLa cell lysate, accessory proteins, reaction mix, nuclease-free water, expression vector and a GFP-positive control vector

Requires:
• 30°C incubator or water bath

Applications:
• Express proteins to measure enzyme activity
• Perform mutational analysis
• Express protein for use in gel mobility shift assays
• Perform co-immunoprecipitation
• Express cytotoxic proteins
• Perform unnatural amino acid labeling

The 1-Step Coupled Human IVT Kit for DNA is a cell-free system using the cellular transcription and translation machinery from a modified HeLa cell line. The procedure is quick and easy and is an effective alternative to other protein expression methods. Simply add an appropriate expression construct to a mixture of HeLa cell lysate, Accessory Protiens, Reaction Mix and incubate at 30°C for 90 minutes to overnight.

The 1-Step Human IVT Kit for mRNA also uses the human protein translation machinery to produce functional protein. This kit is recommended for translation of mRNA transcripts containing an EMCV IRES element and mRNA stabilizing features designed into the pT7CFE1 expression vector.

The human in vitro translation system is robust and will express proteins from a variety of species including mammals, bacteria and protozoa. Benchmarking shows that the expression levels of functional proteins such as luciferase are much higher in the human system compared to either rabbit reticulocyte-based or E. coli-based systems. Furthermore, proteins expressed with the human in vitro translation system are not contaminated with substances that can interfere with downstream applications.

More Product Data
Protocol for in vitro protein expression using mRNA templates
Glycoprotein expression in a human IVT system