Shop All Protein Expression Kits

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

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.

Expi293™ Expression System Kit (Gibco™)

The Expi293™ Expression System is a major advance in transient expression technology for rapid and ultra high-yield protein production from mammalian cells. It is based on high density culture of Expi293F™ cells in Expi293™ Expression Medium. Transient expression is powered by the cationic lipid-based ExpiFectamine™ 293 transfection reagent in combination with specialized transfection enhancers. All components work in concert to generate 2- to 10-fold higher protein yields than previous generation transient expression systems such as the FreeStyle™ 293 Expression System. Expression yields of >1 gram per liter of transfected culture have been demonstrated for some antibody and non-antibody proteins.

The Expi293™ Expression System provides:

Rapid production of milligram to gram quantities of recombinant protein in mammalian cells
High density cultures of Expi293F™ cells that result in more expressing cells per milliliter of culture and higher protein yields
Native folding and mammalian post-translational modifications, such as glycosylation, of expressed proteins
Easy, robust culture and transfection protocols that readily fit into current transient expression workflows
Scalable expression that can be applied to cultures from<1 mL to >10 L, thus generating the amount of protein needed

The components included in the Expi293™ Expression System Kit are: 2 vials of frozen Expi293F™ cells, 1 liter of Expi293™ Expression medium, one ExpiFectamine™ 293 transfection kit sufficient to transfect 1 liter of culture, Opti-MEM® reduced serum media, and an antibody-expressing positive control vector. All components of the Expi293™ Expression System are also available for purchase separately.

Bac-to-Bac™ HBM TOPO™ Cloning Kit (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 (DH10Bac™) rather than homologous recombination in insect cells. The new pFastBac™ HBM TOPO® vector enables secreted protein expression because it has the honeybee melittin (HBM) secretion signal. The new vector should be strongly considered for the expression of glycoproteins. Glycoproteins cannot be glycosylated in the absence of any secretion signal. In contrast to glycoproteins secreted from mammalian cells, glycoproteins secreted from baculovirus can be easily de-glycosylated in vitro. This is a very important feature in order to crystallize proteins!

The pFastBac™ HBM TOPO® vector has also a C-terminal His-Tag with a TEV cleavage signal to enable easy purification of Histidine fusion proteins on nickel-chelating resins (ProBond™ Purification System) and native proteins with the aid of AcTev™ protease.
The vector uses the strong polyhedrin promoter to generate high levels of expression in a variety of insect cell lines such as Sf9, Mimic™ Sf9, Sf21, and High Five™ cells in Sf-900 II & Sf-900™ III media from Gibco.

Traditional baculovirus systems require purification and amplification of an initial low-titer viral supernatant. This requires a time-consuming plaque assay. Bac-to-Bac®'s pFastBac™ vectors, however, recombines with the parent bacmid in DH10Bac™ E. coli competent cells to form an expression bacmid. Transfect the bacmid into insect cells for fast production of a high titer of pure recombinant baculovirus particles in the very first transfection. You'll save weeks of precious time. Collect a pure P2 baculovirus stock on Day 10 without the necessity of tedious plaques assay.

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

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

Champion™ pET161 Gateway™ Expression Kit with Lumio™ Technology (Invitrogen™)

To fit all of your expression needs, Invitrogen offers state-of-the-art Gateway® destination vectors for expression in E. coli, insect, yeast, or mammalian cells, as well as for production of native protein or N- or C-terminal fusion proteins. All Gateway® destination vectors have attR sites for recombination with any attL-flanked fragment, regardless of whether it is an entry clone or an Ultimate™ RF Clone. The following table lists the wide range of destination vectors available.

Additional materials required, available separately: Gateway® entry clone, Gateway® LR Clonase® enzyme mix, and reaction buffer.

Bac-to-Bac™ HBM TOPO™ Secreted 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 HBM TOPO expression system features:

Expression of secreted proteins—The Bac-to-Bac HBM TOPO vector contains the honeybee melittin (HBM) secretion signal to enable secreted protein expression. Because glycoproteins cannot be glycosylated in the absence of a secretion signal, this vector is recommended for the expression of glycoproteins.

Flexibility—The Bac-to-Bac HBM TOPO vector contains a C-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.

Champion™ pET101 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.

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

MembranePro™ Functional Protein Support Kit (Invitrogen™)

For customers studying GPCRs and other membrane proteins, MembranePro™ is the ready-to-use system that delivers enriched, functional membrane proteins efficiently and reliably. Benefits of MembranePro™ include:

• Proteins are displayed on human (or other mammalian) cell membranes
• Cellular quality control and mammalian posttranslational processing helps produce functional protein
• Proteins bud off as lipoparticles and are easily collected from the culture medium
• Lipoparticles are enriched with expressed receptors

MembranePro™ workflow is less labor intensive in comparison to traditional production of membrane fractions. Briefly, the MembranePro™ protocol allows you to:

1. Clone your gene of interest using a TOPO® vector
2. Transfect with Lipofectamine™ 2000 and MembranePro™ Reagent into 293FT cells
3. Harvest lipoparticles 48h post-transfection, concentrate with MembranePro™ Precipitation Mix
4. Store lipoparticles at -80 C or proceed to downstream assays

MembranePro™ products are offered in two configurations – as an expression kit (TOPO® vector, Lipofectamine™ 2000, MembranePro™ Precipitation Mix, MembranePro™ Reagent, 293FT cells) and as a support kit (Lipofectamine™ 2000, MembranePro™ Precipitation Mix, MembranePro™ Reagent). The expression kit is only available in a 10 reaction size; the support kit is available in 10, 60 and 600 reaction sizes.

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.

Champion™ pET102 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®.

Champion™ pET104 BioEase™ Gateway™ Biotinylation System (Invitrogen™)

The Champion™ pET104-DEST BioEase™ Gateway® Expression System provides an easy, efficient method for expressing, purifying, and detecting biotinylated recombinant proteins. The Champion™ pET104-DEST vector (Figure 1) incorporates a 72 amino acid sequence from K. pneumoniae that directs in vivo biotinylation of a specific lysine residue. This tag is recognized and efficiently biotinylated by the enzyme Biotin Protein Ligase (BPL) encoded by the birA gene in E. coli(1). Proteins produced in the Champion™ pET104-DEST vector are expressed as fusion proteins to this sequence. Biotinylated fusion proteins can be purified on streptavidin agarose or detected on western blots using streptavidin-HRP (Figure 2) or streptavidin-AP conjugates.

In addition to the BioEase™ tag, the Champion™ pET104-DEST vector includes the following features:

attR sites for efficient recombination with any attL-flanked Gateway® entry vector
• The T7lac bacteriophage promoter for high-level expression of the recombinant fusion protein
• An enterokinase cleavage site for removal of the N-terminal BioEase™ fusion tag