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

Champion™ pET200 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®.

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

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

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.

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®.

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.

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.

GeneArt™ Cryopreservation Kit for Algae (Invitrogen™)

The GeneArt® Cryopreservation Kit for Algae is used to preserve algal strains and clones for storage at -80°C for years, thus eliminating liquid nitrogen storage and continuous cultures as a way to maintain your clones. Cryopreservation is the optimal choice for preservation and long-term storage of algae, since it minimizes genetic drift, facilitates strain and clone exchange between labs, and helps reduce maintenance labor and costs. Simply grow your cells in the presence of Cryopreservation Reagent A for 2–5 days, harvest, resuspend in Cryopreservation Reagent B, and freeze a small aliquot in one of two recommended freezing containers* at -80°C. The GeneArt® Cryopreservation Kit for Algae has been used to preserve Chlamydomonas and Chlorella strains with 100% resuscitation after thawing.

• Preserve Chlamydomonas and Chlorella strains and clones for -80°C storage
• Eliminate liquid nitrogen storage
• Minimize contamination and genetic drift that typically occur in continuous cultures
• Ship algal clones on dry ice for exchange between labs

To date, most available methods for algae cell preservation have required liquid nitrogen storage, but this method is inconvenient and expensive. Alternatively, researchers have relied on continuous cultures to maintain algal clones, but this often leads to contamination and genetic drift and is not amenable to shipping. The GeneArt® Cryopreservation Kit for Algae provides a way to freeze algae cells at -80°C for long-term storage or shipment on dry ice with minimized loss in viability.

*We have used the following freezing containers with this product:
• Thermo Scientific™ Mr. Frosty® Freezing Container
• Biocision™ CoolCell™ FTS30 Cell Freezing Container

pBAD/TOPO™ ThioFusion™ Expression Kit (Invitrogen™)

The pBAD/TOPO® ThioFusion™ Expression Kit is designed for one-step cloning and regulated expression of thioredoxin fusion proteins in E. coli(Figure 1). The pBAD/Thio-TOPO® vector encodes His-Patch thioredoxin as an N-terminal fusion partner. The thioredoxin fusion can significantly increase the solubility of many difficult-to-express proteins and improve the yield of protein production. The vector includes the following additional features (Figure 2):

• The araBAD promoter for tightly regulated expression in E. coli I-activated vector for 5-minute TOPO® Cloning of Taq-amplified PCR products
• C-terminal polyhistidine (6xHis) tag for purification with nickel-chelating resin and detection with an Anti-His(C-term) Antibody
• C-terminal V5 epitope for detection with an Anti-V5 Antibody
• Enterokinase cleavage site for efficient cleavage of N-terminal fusion tag with EKMax™ Enterokinase

PichiaPink™ Secreted Protein Kit (Invitrogen™)

The PichiaPink™ Secreted Protein Expression Kit is a new recombinant protein expression system based on the yeast Pichia pastoris. The PichiaPink™ Secreted Protein Expression Kit gives you convenient and cost-efficient bioproduction from shake flask to 1000-liter production of your recombinant protein. The PichiaPink™ Secreted Protein Expression Kit comes with our new Pichia strains, the pPINK-HC and pPINK-LC vectors, and secretion signal sequences for production of your protein.
  Get started with secreted protein production in Pichia pastoris.
•   Screen your Pichia transformants rapidly by auxotrophy and color selection.
•   Reduce protein degradation with protease-deficient Pichia pastoris strains.

The PichiaPink™ Secreted Protein Expression Kit comes with:
•   PichiaPink™ Secreted Protein Vector Kit (Cat. No. A11153)
•   PichiaPink™ Expression Strain Set (Cat. No. A11154)
•   PichiaPink™ Media Kit (Cat. No. A11156)

A New Way to Screen Pichia Transformants
The PichiaPink™ Secreted Protein Expression Kit uses adenine auxotrophy to select for Pichia transformants. The PichiaPink™ strains have the ade2 genotype and require complementation with the ADE2 gene to grow without adenine. Use of an auxotrophic marker also makes it easier to maintain transformants. No antibiotics are required to maintain your transformants.

In addition, you can screen for Pichia transformants by color. The untransformed PichiaPink™ strains appear as pink colonies while transformed PichiaPink™ strains appear as white colonies (Fig 1).

Secrete Your Proteins with the pPINKα-HC Vector

The PichiaPink™ Secreted Protein Expression Kit comes with the pPINKα-HC vector. The pPINKα-HC vector has the α-mating factor signal sequence built-in and allows your protein to be secreted into the media. Secreting your protein into the media makes harvesting and purifying your protein easier. The pPINKα-HC vector is built around the ADE2 gene for complementing the ade2-deficient Pichia strains. This vector uses the methanol-induced AOX1 promoter to express your protein. Further, the pPINKα-HC vector is designed to be integrated at high-copy-number in your Pichia transformants.

The PichiaPink™ Secreted Protein Vector Kit (Cat. No. A11153) can be ordered separately to refill the PichiaPink™ Secreted Protein Expression Kit.

Keep Your Proteins Intact
The PichiaPink™ Secreted Protein Expression Kit comes with the PichiaPink™ Expression Strain Set. This set contains the 4 Pichia pastoris strains for use with the PichiaPink™ Yeast Expression System. All 4 strains have the ade2 genotype. In addition, 3 strains include knockouts in 2 protease genes. Protease-deficient strains reduce the need for protease inhibitors when growing your cells and improve the yield of your protein. Test a single protease knockout or the double protease knockout to find the one that works best for you.

You can order the PichiaPink™ Expression Strain Set (Cat. No. A11154) separately to refill the PichiaPink™ Secreted Protein Expression Kit .

Media Pouches to Get You Started

The PichiaPink™ Media Kit is a set of convenient prepackaged media pouches for use with the PichiaPink™ Secreted Protein Expression Kit. The PichiaPink™ Media Kit contains media needed to grow PichiaPink™ strains from frozen stocks to cultures ready for transformation and selection. You can order the PichiaPink™ Media Kit (Cat. No. A11156) separately to refill the PichiaPink™ Secreted Protein Expression Kit.

Why Choose the PichiaPink™ Yeast Expression System?
The PichiaPink™ Yeast Expression System is based on the yeast Pichia pastoris. Advantages of Pichia pastoris include rapid growth, well-defined genetic background, simple media formulation, and easy handling. For over 30 years, Pichia pastoris has been used by labs around the world for producing hundreds of different proteins from many species including human (Ref 1, 2). The PichiaPink™ Yeast Expression System allows convenient and cost-effective protein production from small to large scales.

For information on obtaining a commercial-use license for the PichiaPink™Yeast Expression System, please inquire at outlicensing@lifetech.com.

For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use.

Related Links
Learn more about the PichiaPink™ Yeast Expression System.
Learn more about our other protein expression systems.

References

1. Cereghino JL, Cregg JM. Heterologous protein expression in the methylotrophic yeast Pichia pastoris. FEMS Microbiol Rev. 2000 Jan;24(1):45-66. [PubMed]
2. Cereghino GP, Cereghino JL, Ilgen C, Cregg JM. Production of recombinant proteins in fermenter cultures of the yeast Pichia pastoris. Curr Opin Biotechnol. 2002 Aug;13(4):329-32. [PubMed]

MAX Efficiency™ Transformation Reagent for Algae (Invitrogen™)

MAX Efficiency® Transformation Reagent for Algae is a buffer that enhances transformation efficiency for multiple strains of Chlamydomonas species. One of the biggest hurdles in research and development with Chlamydomonas has been the introduction of exogenous DNA into Chlamydomonas strains due to rigid cell walls. Methods such as glass bead agitation, electroporation, and microparticle bombardment are available but often result in low transformation efficiency.

MAX Efficiency® Transformation Reagent for Algae increases the permeability of the Chlamydomonas cell wall and facilitates increased delivery of DNA into the cell nucleus by electroporation. Simply grow the Chlamydomonas cells to early log phase, harvest by centrifugation, and wash twice with MAX Efficiency® Transformation Reagent prior to electroporation. To date, we have seen >200-fold increase in transformation efficiency over previously recommended conditions in 10 different Chlamydomonas strains, including wild type and mutants, using circular or linear DNA, as well as PCR fragments.

• Obtain >1000 transformants/µg DNA for cell wall(+) strains
• Obtain >100 transformants/µg DNA for cell wall(-) strains
• Transform circular or linear DNA or PCR fragments

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.

Expressway™ Maxi Cell-Free E. coli Expression System (Invitrogen™)

The Expressway™ Maxi Cell-Free E. coli Expression System uses an efficient, coupled transcription/translation reaction to produce milligram quantities of soluble, functionally active protein in 4-6 hours. The procedure can be performed in a single reaction tube and is easily scalable without the need for specialized equipment. The TOPO TA Cloning® expression vectors (5-min cloning with 95% efficiency) are provided for optimal expression results. In addition to all the advantages of an open expression system, Expressway™ Maxi technology provides a means to produce high levels of recombinant protein that may be easily detected and purified for various downstream applications.

Milligram level protein production within 4-6 hours
The key to the Expressway™ Maxi technology is the unique formulation of the Feed Buffer that boosts the reaction productivity so that more than one milligram of protein is produced in a 2-ml reaction within 4-6 hours (Figure 1). The formulation of the lysate enables protein synthesis using either circular or linear templates.

High-throughput (HTP) compatible
This system is also designed for HTP expression. One kit is good for 200 x 50-reactions, which can be accommodated by 2 x 96-well plates. Without feed buffer, such reactions need only two hour’s time. Once a positive expression is detected, scale-up protein production can be performed with the large Expressway™ cell-free format or in E. coli cell-based systems.

Optimized vectors
pEXP5 TOPO® TA vectors are optimized for use in the Expressway™ Milligram system (Figure 2). They are designed to minimize additional amino acid sequences that may interfere with native protein folding and functionality. Features of these vectors include:

• pEXP5-NT/TOPO® expression vector provides N-terminal histidine tag and TEV protease recognition site

• pEXP5-CT/TOPO® expression vector can be used for expressing proteins with a C-terminal histidine tag or for native protein expression by introducing a stop codon at the end of your gene of interest

Simple and fast procedure
The Expressway™ Maxi Cell-Free Expression System provides all the components needed for optimal cell-free protein production. The system includes an E. coli extract, IVPS reaction buffer, Feed Buffer, T7 enzyme mix, 19 amino acid mix, and individual tubes of methionine. To start a 2-ml reaction, mix the reaction buffer, 19 amino acid mix, your choice of labeled or unlabeled methionine and cysteine, T7 enzyme mix, and your DNA template (with T7 promoter) with the E. coli extract. After a 30-minute incubation, add the Feed Buffer. Milligram-levels of active protein will be synthesized within 4-6 hours. To perform HTP expression, simply premix all the component and aliquot into a HTP format.