Shop All Protein Expression Kits

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

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

GeneArt™ Synechococcus Protein Expression Vector (Invitrogen™)

The GeneArt™ Synechococcus Protein Expression Vector is our second-generation Synechococcus protein expression vector optimized for relative high-level expression in algae and with dual protein tags for detection and/or purification of your gene of interest. The kit includes expression vector and easy-to-follow protocols. Our Gibco™ BG-11 Media, sold separately, is optimized for the growth and maintenance of select Cyanobacteria including Synechococcus elongatus, available from the ATCC collection.

• Express >10% total soluble protein of your gene of interest
• Detect and purify your gene of interest with 6His-TEV and/or V5-His epitope tags
• Compatible with seamless assembly for creation of constructs

A specific decided expression vector
The GeneArt Synechococcus Protein Expression Vector is designed to provide constitutive expression in Synechococcus elongatus. Expression is controlled by promoter psbA1, a ribosome binding site (RBS), and optional initiation and stop codons depending on gene location. The vector provides purification and detection tags at the N- and C-termini: an N-terminal polyhistidine tag with a TEV recognition site for optional cleavage and a C-terminal V5 epitope followed by a polyhistidine tag. Two MCS sites provide the flexibility to attach either, both, or no tags toyour protein. Our pSyn_6 vector is also compatible with seamless cloning, an optional cloning method that results in no extra sequences in your final construct.

Features of the vector include:
• Strong constitutive promoter, psbA1, for robust expression of recombinant gene of interest
• Ribosome binding site (RBS) for improved expression of some genes
• Targeted integration of your gene of interest into the Synechococcus elongatus genome
• N-terminal 6His-TEV and C-terminal V5-6His epitope tags
• Spectinomycin-resistance gene for selection in Synechococcus
• Flexible, multiple cloning site vector

>80% integration efficiency
The transformation of Synechococcus elongatus PCC7942 relies on homologous recombination between the cell’s chromosome and exogenous DNA that is not autonomously replicating and contains sequences homologous to the chromosome. The location of integration into the chromosome (neutral site, NS1) was developed as a cloning locus (Clerico et al., 2007), since it can be disrupted without any aberrant phenotype, thus allowing recombination of ectopic sequences. When transformed with vectors containing an antibiotic resistance cassette and neutral site sequences, a double homologous recombination event occurs between the neutral site vector and the Synechococcus elongatus chromosome. The selective marker (spectinomycin) and the gene of interest driven by a promoter are inserted into the neutral site and the vector backbone (pUC) is lost, allowing the expression of recombinant genes in Synechococcus elongatus PCC 7942.

Gibco BG-11 Media—optimized for cyanobacteria
Gibco BG-11 Media, available separately, is optimized for the growth and maintenance of select Cyanobacteria including Synechococcus elongatus. The 1X formulation lets you avoid laborious media preparation steps. Award-winning bottle design is easier to use in the biosafety cabinet, minimizes the risk of contamination, and helps you perform cell culture more consistently. Superior packaging and quality, greater reliability, and improved consistency in Cyanobacteria culture results in better overall efficiency and more robust data.

GeneArt™ Chlamydomonas Protein Expression Vector (Invitrogen™)

The GeneArt™ Chlamydomonas Protein Expression Vector is our second-generation Chlamydomonas protein expression vector for transgene expression from the nuclear genome of eukaryotic green alga Chlamydomonas reinhardtii 137c. It is optimized for relative high-level expression, provides selection against gene silencing, and offers dual protein tags for detection and/or purification of your gene of interest. The kit includes expression vector and easy-to-follow protocols. Our Gibco™ TAP Media, offered separately, is optimized for the growth and maintenance of Chlamydomonas. Chlamydomonas reinhardtii 137c is available from the Chlamydomonas Resource Center.

• Express up to 1% total soluble protein of your gene of interest
• Select against gene silencing even over multiple passages
• Detect and purify your gene of interest with 6His-TEV and/or V5-His epitope tags
• Compatible with seamless assembly for creation of your constructs
• Enables you to receive reliable results with exceptional strain viability and purity

Chlamydomonas pChlamy_4 vector
Transgene expression from the Chlamydomonas nuclear genome offers several advantages over chloroplast expression, such as post-translational modifications and protein-targeting and/or secretion. However, expression from the nuclear genome has typically been less than robust, and the molecular mechanism(s) of poor expression are not completely understood. Possible reasons include poor promoters, genome integration position effects, and transgene silencing. The GeneArt Chlamydomonas Protein Expression Vector provides relative high levels of expressed protein and selection against silencing. Our pChlamy_4 vector is also compatible with seamless cloning, an optional cloning method that results in no extra sequences in your final construct.

Features of the vector include:
• Endogenous and constitutive promoter, Hsp70A-RbcS2, is composed of activator Hsp70A and two copies of RBC S2 intron sequence, resulting in increased expression of your gene of interest
• Hsp70A-Rbc S2 hybrid promoter fused to the bleomycin/Zeocin™-resistance gene allows positive transformants to maintain protein expression levels for multiple passages
• Self-cleaving sequence for the 2A peptide from foot-and-mouth-disease-virus (FMDV) mediates proper cleavage between resistance markers and protein-of-interest to yield two discrete proteins
• 3’ UTR for proper transcript termination and possible additional benefits like increased translation efficiency, mRNA stability, and polyadenylation signals
• Dual protein tags 6His-TEV and V5-His epitopes can be fused to both or either ends of your gene of interest or no tag at all

Selection against silencing
In order to circumvent the transgene silencing that often occurs in C. reinhardtii, our new pChlamy_4 vector is designed so that proteins are expressed as transcriptional fusions with the bleomycin/zeocin resistance gene sh-ble (Rasala, et al, 2012). The self-cleaving sequence for the 2A peptide from the foot-and-mouth-disease-virus (FMDV) is placed between the antibiotic resistance gene and the gene of interest. It encodes a short ~20 amino acid sequence that mediates proper cleavage to yield two discrete proteins. With this system we have seen positive transformants maintain high expression levels for much longer than with other systems, even after many passages with or without selection pressure.

MAX efficiency transformation reagent for algae
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, when used to pretreat the cells prior to electroporation, enhances transformation efficiency for multiple strains of Chlamydomonas species. It increases the permeability of the Chlamydomonas cell wall and facilitates increased delivery of DNA into the cell nucleus by electroporation. To date, we have seen >200 fold increase in transformation efficiency over previously recommended transformation conditions in 10 different Chlamydomonas strains, including wild type and mutants, using circular or linear DNA, as well as PCR fragments.

Gibco TAP Media—optimized for Chlamydomonas
Gibco TAP Media, available separately, is optimized for the growth and maintenance of Chlamydomonas. The 1X formulation lets you avoid laborious media preparation steps. Award-winning bottle design is easier to use in the biosafety cabinet, minimizes the risk of contamination, and helps you perform cell culture more consistently. Superior packaging and quality, greater reliability, and improved consistency in Chlamydomonas culture results in better overall efficiency and more robust data.

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

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

Freedom™ CHO-S™ Kit (Gibco™)

Gibco® Freedom® CHO-S® Kit, co-developed with ProBioGen AG, is an easy-to-use, beginning-to-end product, for cloning and expression of recombinant proteins in Chinese Hamster Ovary (CHO)-derived suspension culture. The Gibco® Freedom® CHO-S® Kit includes components for transfection, expression, clone creation, and stable cell line selection, all in a conveniently packaged kit. The Gibco® Freedom® CHO-S® Kit also delivers high titers, and offers simplified commercial licensing options.

• Flexible commercial licensing without the need for royalties
• Ability to achieve IgG titers over 3g/L
• Complete workflow that can take your gene of interest from transfection to lead clone typically in 5 months
• Convenient packaging, for simplified ordering and storage

Commercial Licensing that is Simple, Flexible and Without Royalties
Multiple companies provide CHO (Chinese Hamster Ovary) stable cell line development services and products that require annual fees during development of your therapeutic protein. In addition, royalties may be required on the sales of your therapeutic protein upon commercialization. Gibco® Freedom® Kits provide you with the tools that enable you or your CRO provider to develop your stable cell line for just the price of the kit itself. Furthermore, as you move towards commercial use, a flexible license structure is available without the burden of royalties and works to keep it as simple as a onetime payment (Fig 1). For more information regarding licensing, email us at outlicensing@lifetech.com.

Industry Standard Production Levels
Current bioprocess industry standards to achieve manufacturing efficiencies for IgG type molecule production with stable CHO cell lines in bioreactors is 2-3g/L or better. We have demonstrated that the Gibco® Freedom® CHO-S® Kit can consistently generate IgG producing scalable clones (stable out to 60 population doublings), that meet this standard of productivity (Fig 2).

Complete, Fast, and Efficient Workflow
The Gibco® Freedom® CHO-S® Kit’s complete, proven, and fully integrated workflow allows you to move from transfection to lead clone typically in 5 months, with reduced hands on time, and reduced Fulltime Equivalent (FTE) requirements (Fig 3). The entire workflow is performed in our CD FortiCHO™ medium, a chemically defined, animal origin free (AOF) medium. The kit also offers the following advantages:

• Cloning of one or two genes that encode your protein(s) of interest
• High efficiency transfection of DNA into CHO-S® Cells (cGMP-banked)
• Included flash drive with loaded manual

New Convenient Packaging
To free you to focus on the creation of your stable cell line rather than on figuring out where and when all the components of your experiment are arriving, Freedom® CHO-S® Kit introduces the smART™ tote packaging system (shown in above product photos). All components for the creation of your expression cell line are delivered in soft cooler totes along with a parts flyer, and a flash drive containing the manual. Upon delivery you have all the information needed for correct storage and use of components consolidated in the two smART™ tote shipment coolers.

Access to Gibco® Freedom® Kit Support
If any specific support is needed beyond the manual, please contact your account manager to obtain a Process Science Manager consultation. In addition, one of our support scientists may check in with you to ensure that your experience with the kit is as successful as possible. For further assistance, please email our support team at GibcoServices@thermofisher.com.

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

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 (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.

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.

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®.

Expi293™ MembranePro™ Expression System (Gibco™)

For researchers studying G protein–coupled receptors (GPCRs) and other membrane proteins, the Expi293™ MembranePro™ Expression System delivers enriched, functional membrane proteins with high efficiency, scalability and reliability for a wide variety of low or high throughput applications.

Features of the Expi293™ MembranePro™ Expression System include:
• Efficient—more than 20-fold increase in membrane protein yield compared to the adherent culture system
• Scalable—easy scale-up without the need for multiple flasks
• Easy to use—no enzymatic or mechanical dissociation required

In addition, benefits of MembranePro™ technology include:
• Proteins are displayed on human (or other mammalian) cell membranes
• Cellular quality control and mammalian post-translational processing helps produce functional protein
• Proteins bud off as lipoparticles and are easily collected from the culture medium
• Lipoparticles are enriched with expressed receptors

With the less labor-intensive Expi293™ MembranePro™ Expression System protocol, you will:
1. Clone your gene of interest using a TOPO® vector
2. Transfect a susupension culture of Expi293F™ cells using Expifectamine™ 293 transfection reagent
3. Harvest lipoparticles 48 hrs after addition of enhancers using MembranePro™ Precipitation Mix
4. Store lipoparticles at -80 C or proceed to downstream assays

Note: Exi293F™ Cells (A14527) and a pEF6 V5-His TOPO® Expression Kit (K9610-20) are required for use of this system and must be purchased separately when needed.

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]

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

Champion™ pET160 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.