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.

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

Expressway™Lumio™ Expression and Detection System, without vector Invitrogen™

The Expressway™ Lumio™ Expression and Detection System takes advantage of the Lumio™ recognition sequence, a small, six amino acid sequence (Cys-Cys-Pro-Gly-Cys-Cys). The Lumio™ detection reagent binds the recognition sequence with high specificity and affinity, resulting in a bright fluorescent signal for real-time protein production analysis and immediate in-gel protein detection. In addition, Expressway’s specialized E. coli lysate, derived from a slyD mutant, eliminates nonspecific binding of the Lumio™ Green Detection Reagent to the endogenous SlyD protein and provides optimal background for detection of recombinant proteins (Figure 1). The Lumio™ Green Detection Kit is included in the system for real-time detection of protein synthesis as well as easy in-gel protein detection. The Lumio™ vectors (Figures 2 and 3) also feature:

attR sites for efficient recombination with any attL-flanked Gateway® entry vector
N-terminal Lumio™ tag (pEXP3-DEST vector) with TEV cleavage site for efficient removal of the Lumio™ sequence following purification
C-terminal Lumio™ tag (pEXP4-DEST vector) for easy, and immediate in-gel detection
T7 promoter, ribosome binding site, and T7 terminator optimally spaced for cell-free protein expression

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®.

Expi293™ Inducible GnTI- Expression System Kit Gibco™

The Expi293 Inducible GnTI- Expression System Kit is part of the Expi293 platform for rapid, high-yield transient protein expression from mammalian 293 cells. The system kit is centered on engineered Expi293F cells that do not have N-acetylglucosaminyltransferase I (GnTI) activity, therefore lacking complex N-glycans, and stably express the tetracycline repressor (TetR) protein, thus enabling tetracycline-regulated protein expression. The Expi293F Inducible GnTI- cell line was established by the selective knockout of the GnTI gene and the stable integration of the regulatory plasmid pcDNA6/TR, which encodes the TetR gene under the control of the human CMV promoter, in parental Expi293F cells. This cell line is a powerful tool for tetracycline-inducible high-yield expression of homogeneously glycosylated recombinant proteins.

Expi293 Inducible GnTI- Expression System Kit features include:
• Homogeneous N-glycosylation of expressed proteins
• Tetracycline-inducible protein expression with extremely low basal expression levels
• Equivalent protein yields to parental Expi293F cells—up to 1 g/L
• pcDNA5/TO expression vector for tetracycline-regulated expression of the gene of interest
• Suspension, high-density culture of Expi293F Inducible GnTI- cells in Expi293 Expression Medium
• High-efficiency transient transfection using ExpiFectamine 293 reagent in combination with specialized transfection enhancers

The components included in the Expi293 Inducible GnTI- Expression System Kit are: one vial of frozen Expi293F Inducible GnTI- cells, 1 L of Expi293 Expression Medium, one ExpiFectamine 293 transfection kit sufficient to transfect 1 L of culture, 100 mL Opti-Plex Complexation Buffer, one PNGase F Glycan Cleavage kit, one pcDNA5/TO Mammalian Expression Vector kit, 500 mg tetracycline hydrochloride, and the pRABBIT IgG IRES-EmGFP positive control vector. All components of the Expi293 Inducible GnTI- Expression System are also available for individual purchase.

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 Proteins, 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

Expressway™ Maxi Cell-Free E. coli Expression System with pEXP5-NT/TOPO™ and pEXP5-CT/TOPO™ vectors 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.

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.

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

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.

Expi293™ GnTI- Expression System Kit Gibco™

The Expi293 GnTI- Expression System Kit is part of the Expi293 platform for rapid, high-yield transient protein expression from mammalian 293 cells. The system kit is centered on engineered Expi293F cells that do not have N-acetylglucosaminyltransferase I (GnTI) activity and therefore lack complex N-glycans. The Expi293F GnTI- cell line was established by the selective knockout of the GnTI gene in parental Expi293F cells. This cell line is a powerful tool for the high-yield expression of homogeneously glycosylated recombinant proteins.

Expi293 GnTI- Expression System Kit features include:
• Homogeneous N-glycosylation of expressed proteins
• Equivalent protein yields to parental Expi293F cells—up to 1 g/L
• Significantly higher protein yields than other HEK293 GnTI- cell line alternatives
• Suspension, high-density culture of Expi293F GnTI- cells in Expi293 Expression Medium
• High-efficiency transient transfection using ExpiFectamine 293 reagent in combination with specialized transfection enhancers

The components included in the Expi293 GnTI- Expression System Kit are: one vial of frozen Expi293F GnTI- cells, 1 L of Expi293 Expression Medium, one ExpiFectamine 293 transfection kit sufficient to transfect 1 L of culture, 100 mL Opti-Plex Complexation Buffer, one PNGase F Glycan Cleavage kit, and the pRABBIT IgG IRES-EmGFP positive control vector. All components of the Expi293 GnTI- Expression System are also available for individual purchase.

MembranePro™ Functional Protein Expression 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.

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.

Expi293™ Met (–) Protein Labeling Kit Gibco™

The Expi293 Met (–) Protein Labeling Kit includes Expi293 Met (–) Expression Medium and the ExpiFectamine 293 Met (–) Transfection Kit for rapid, high-yield transient protein expression from Expi293F cells for applications where labeling of methionine residues in proteins is desired. The kit components are specifically formulated without methionine, allowing users to add labeled methionine of their choice.

Expi293 Met (–) Protein Labeling Kit features include:
• Methionine-free medium and transfection enhancer formulations
• 2- to 10-fold higher protein yields compared to other transfection reagents used for high-density 293 cell culture
• Same transient transfection protocol as ExpiFectamine 293 Transfection Kit
• Scalable transfection from small to large scale while maintaining equivalent volumetric protein yields

The Expi293 Met (–) Protein Labeling Kit is specifically designed to easily integrate with other components of the Expi293 Expression System, including Expi293F cells and Expi293 Expression Medium. The components included in the Expi293 Met (–) Protein Labeling Kit are: 1 L Expi293 Met (–) Expression Medium and one ExpiFectamine 293 Met (–) Transfection Kit sufficient to transfect 1 L of culture.

L-methionine (methyl-13C) and L-selenomethionine labeling reagents are available separately.

ExpiSf™ Protein Production Kit Gibco™

The ExpiSf Protein Production Kit is designed to significantly boost protein expression from high-density suspension cultures of Sf9 insect cells. The kit includes the chemically defined, animal origin-free ExpiSf Enhancer and ExpiSf CD Medium, which together power the highest possible level of protein expression from baculovirus-infected ExpiSf9 Cells.

The ExpiSf Protein Production Kit:
• Delivers greater than 80% more protein yield in high-density suspension Sf9 cell culture than ExpiSf CD Medium alone
• Includes ExpiSf Enhancer, which is specifically designed to work in concert with ExpiSf CD Medium to significantly boost protein expression from baculovirus-infected insect cell cultures
• Demonstrates direct scalability of baculovirus infection from culture volumes of 4 mL in deep-well plates to >3 L in shake flasks

The ExpiSf Protein Production Kit is specifically designed to work with ExpiSf9 Cells in the ExpiSf Expression System. The kit supplies sufficient ExpiSf Enhancer and ExpiSf CD Medium to infect 1 L, 10 L, or 50 L of culture. ExpiSf CD Medium is also available for purchase separately.

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