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TC-FlAsH™ TC-ReAsH™ II In-Cell Tetracysteine Tag Detection Kit, with Mammalian TC-Tag Gateway™ Expression Vectors (Green Fluorescence) (Red Fluorescence) (Invitrogen™)

TC-FlAsH™ TC-ReAsH™ II In-Cell Tetracysteine Tag Detection Kit is an expression tag-based fluorescence labeling system for live-cell labeling. The expression construct is created when the Gateway entry clone (containing the gene of interest) is recombined with one of two destination vectors (pcDNA6.2/nTC-Tag-DEST for N-terminal tagging or pcDNA6.2/cTC-Tag-DEST for C-terminal tagging). The expression construct is then used to transform the host strain. The protein can be detected either with green-fluorescent FlAsH-EDT2 reagent or red-fluorescent ReAsH-EDT2 reagent. The tag is very small (6 amino acids) and the protein of interest is fluorescent only when the labeling reagent is added. BAL wash buffer replaces the previously supplied Disperse Blue 3 and EDT-based wash buffer as an olfactorily more agreeable reagent that yields superior signal to noise.

The kit contains FlAsH-EDT2 labeling reagent, ReAsH-EDT2 labeling reagent, BAL wash buffer, pcDNA6.2/cTC-Tag-DEST, pcDNA6.2/nTC-Tag-DEST, and pcDNA6.2/nTC-Tag-p64 Control Plasmid. BAL wash buffer should be stored at 4°C, and the rest of the kit stored at -20°C, protected from light. The kit is stable for 6 months when stored correctly.

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

pTrcHis A, B, & C Bacterial Expression Vectors (Invitrogen™)

Our pTrcHis A, B, & C vectors are designed to offer enhanced translation initiation and high-level expression in E. coli. These vectors feature:

• High-level regulated transcription from the trc promoter
• Enhanced translation efficiency of eukaryotic genes in E. coli
• The lacO operator and lacIq repressor gene for transcriptional regulation in any E. coli strain

This particular vector offers:

• N-terminal polyhistidine (6xHis) tag for rapid purification with nickel-chelating resin and detection with an anti-HisG antibody
• N-terminal Xpress™ epitope for easy detection with an anti-Xpress™ antibody
• Enterokinase cleavage site for removal of fusion tag

For C-terminal polyhistidine tag and c-myc epitope, please see our pTrcHis2 A, B, & C Vector.

pYES2/NT A, B, & C Yeast Expression Vectors (Invitrogen™)

pYES2/NT and pYES2/CT are S. cerevisiae expression vectors derived from the parental pYES2 vector. Both vectors offer the URA3 gene for selection in yeast. pYES2/NT features an N-terminal Xpress™ epitope for detection with Invitrogen's Anti-Xpress™ Antibody and a polyhistidine (6xHis) tag for purification with nickel-chelating resin. pYES2/CT contains a C-terminal V5 epitope for detection and a polyhistidine (6xHis) tag for purification.

pcDNA™6/myc-His A, B, & C Mammalian Expression Vectors (Invitrogen™)

All pcDNA™ vectors contain a strong promoter for high-level expression in mammalian cells, a choice of selection marker for generating stable cell lines, and an epitope tag for easy detection with a monoclonal antibody and rapid purification on nickel-chelating resin. Each vector is available in three reading frames to simplify cloning in-frame with the fusion tag.

pcDNA™6/V5-His A, B, & C Mammalian Expression Vectors (Invitrogen™)

All pcDNA™ vectors contain a strong promoter for high-level expression in mammalian cells, a choice of selection marker for generating stable cell lines, and an epitope tag for easy detection with a monoclonal antibody and rapid purification on nickel-chelating resin. Each vector is available in three reading frames to simplify cloning in-frame with the fusion tag.

pGAPZα A, B, & C Pichia pastoris Expression Vectors (Invitrogen™)

pGAPZ A, B, & C and pGAPZ A, B, & C are expression vectors designed for high-level, constitutive expression in Pichia pastoris. pGAPZ and pGAPZ were created by replacing the methanol-regulated AOX1 promoter with the constitutive, glyceraldehyde-3-phosphate dehydrogenase (GAP) promoter in the backbone of the pPICZ vectors. Although the yield
of any protein constitutively expressed in the Pichia system is dependent on the toxicity of the protein to yeast, constitutive expression under the control of pGAPZ or pGAPZ vectors can sometimes produce greater yields than inducible expression (1).

pTrcHis2 A, B, & C Bacterial Expression Vectors (Invitrogen™)

Our pTrcHis A, B, & C vectors are designed to offer enhanced translation initiation and high-level expression in E. coli. These vectors feature:

• High-level regulated transcription from the trc promoter
• Enhanced translation efficiency of eukaryotic genes in E. coli
• The lacO operator and lacIq repressor gene for transcriptional regulation in any E. coli strain

This particular vector offers:

• C-terminal polyhistidine (6xHis) tag for rapid purification with nickel-chelating resin and detection with an anti-His(C-term) antibody
• C-terminal c-myc epitope for easy detection of fusion proteins with an anti-myc antibody

For N-terminal polyhistidine tag and Xpress™ epitope, please see our pTrcHis A, B, & C Vector.

pcDNA™4/HisMax A, B, & C Mammalian Expression Vectors (Invitrogen™)

pcDNA™4/HisMax is specifically designed to maximize protein expression in a variety of mammalian cells. The vector contains the QBI SP163 translational enhancer to increase expression levels two- to five-fold above those seen with the promoter alone (Figure 1). In addition to SP163-enhanced expression, pcDNA™4/HisMax includes a cleavable N-terminal Xpress™ tag for rapid detection of recombinant protein with an Anti-Xpress™ Antibody (Figure 2). pcDNA™4/HisMax is available TOPO® Cloning-ready for five-minute cloning of Taqamplified PCR products.

pMT/BiP/V5-His A, B, & C Drosophila Expression Vectors (Invitrogen™)

The DES®-Inducible/Secreted Kit provides the vector pMT/BiP/V5-His for inducible, secreted expression of recombinant proteins. This vector offers the inducible metallothionein promoter that is induced upon addition of copper sulfate or cadmium chloride. The N-terminal signal sequence from the insect BiP gene is provided to direct the recombinant fusion protein through the secretory pathway of S2 cells into the culture medium. The pMT/BiP/V5-His vector offers the following additional features:

• Small size (3.6 kb) to improve DNA yields and increase subcloning efficiency
• C-terminal V5 epitope tag for rapid detection with Invitrogens Anti-V5 Antibody
• C-terminal 6xHis tag for simple purification of recombinant fusion proteins using nickel-chelating resin

To facilitate cloning, a set of three vectors-A, B, and C-is provided. Each vector has the multiple cloning site in a different reading frame relative to the BiP signal sequence.

pcDNA™4/TO/myc-His A, B, & C Mammalian Expression Vectors (Invitrogen™)

A Tetracycline-Regulated Expression System without Viral Transactivators
The T-REx™ System yields higher levels of induced expression than any other regulated mammalian expression system. It utilizes the complete CMV promoter and adds control elements from the bacterial tetracycline resistance operon to effectively repress and derepress transcription from one of the strongest mammalian promoter sequences known (1,2).

Specific Activation
The T-REx™ System uses a repressor mechanism that blocks transcription from the powerful CMV promoter in the absence of tetracycline. Because the T-REx™ System elements do not use viral transactivators, you can achieve high-level expression from the complete CMV promoter without secondary, non-specific activation of host genes.

The T-REx™ Mechanism
The T-REx™ transcriptional control elements are illustrated in Figure 1. Two tetracycline operator sequences (TetO2) have been inserted between the TATA box of the CMV promoter and the transcriptional start site. The TetO2 sequence itself has no effect on expression. When the tetracycline repressor protein (TR) is present, it effectively binds the TetO2 sites and blocks transcription initiation. Tetracycline added to the culture medium binds to, and changes the conformation of, the TR protein. This change causes the TR protein to release the TetO2 sites, derepressing transcription from the CMV promoter. The result is high-level expression of the gene of interest (Figure 2). Expression levels can be modulated based on the tetracycline concentration and can be induced to levels that are achieved with constitutive CMV expression vectors.
T-REx™ is a powerful inducible mammalian expression system that allows you to regulate expression from the complete human cytomegalovirus (CMV) enhancer-promoter. T-REx™inducible expression vectors offer the following features:

• Complete CMV enhancer-promoter sequence containing two copies of the tetracycline operator TetO2 sequence for high-level regulated expression
• Zeocin™ or hygromycin resistance gene for effective selection of stable mammalian cell lines
• Large multiple cloning site to simplify cloning

In addition, pcDNA™4/TO/myc-His offers a c-myc epitope for rapid detection of the recombinant protein with an Anti-myc Antibody and a polyhistidine (6xHis) sequence for simple purification of the recombinant protein with nickel-chelating resin and detection with Anti- His(C-term) Antibody.

The regulatory vector, pcDNA™6/TR, is provided for high-level expression of the tetracycline repressor (TR) protein. This vector expresses the Blasticidin resistance gene for rapid selection of mammalian cell lines that stably express the TR protein.

pEF1/His A, B, & C Mammalian Expression Vectors (Invitrogen™)

All pEF or pUB vectors contain a strong promoter for high-level expression in mammalian cells, a choice of selection marker for generating stable cell lines, and an epitope tag for easy detection with a monoclonal antibody and rapid purification on nickel-chelating resin. Each vector is available in three reading frames to simplify cloning in-frame with the fusion tag.

pPIC6 A, B, & C Pichia Vectors (Invitrogen™)

pPIC6 A, B, & C are Pichia pastoris expression vectors designed for simple selection,
high-level expression, and rapid protein purification and detection. The pPIC6 A,
B, & C vectors are designed for high-level secreted expression. Both sets of vectors
have the following features:
Blasticidin resistance for direct selection of multi-copy integrants
Inducible AOX1 promoter for high-level expression in Pichia pastoris
C-terminal c-myc epitope for convenient detection with an Anti-myc Antibody
C-terminal polyhistidine (6xHis) tag for rapid purification with nickel-chelating
resin and detection with an Anti-His(C-term) Antibody
pPIC6 also features:
An É -factor secretion signal for efficient transport of proteins to the medium

pcDNA™3.1/His A, B, & C Mammalian Expression Vectors (Invitrogen™)

All pcDNA™ vectors contain a strong promoter for high-level expression in mammalian cells, a choice of selection marker for generating stable cell lines, and an epitope tag for easy detection with a monoclonal antibody and rapid purification on nickel-chelating resin. Each vector is available in three reading frames to simplify cloning in-frame with the fusion tag.

pUB6/V5-His A, B, & C Mammalian Expression Vectors (Invitrogen™)

All pEF or pUB vectors contain a strong promoter for high-level expression in mammalian cells, a choice of selection marker for generating stable cell lines, and an epitope tag for easy detection with a monoclonal antibody and rapid purification on nickel-chelating resin. Each vector is available in three reading frames to simplify cloning in-frame with the fusion tag.

pEF1/V5-His A, B, & C Mammalian Expression Vectors (Invitrogen™)

All pEF or pUB vectors contain a strong promoter for high-level expression in mammalian cells, a choice of selection marker for generating stable cell lines, and an epitope tag for easy detection with a monoclonal antibody and rapid purification on nickel-chelating resin. Each vector is available in three reading frames to simplify cloning in-frame with the fusion tag.

Mammalian Lumio™ Gateway™ Vectors, with Lumio™ Green In-Cell Detection Kit (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.

pTracer™-EF A, B, & C Mammalian Expression Vectors (Invitrogen™)

Three pTracer™ mammalian expression vectors are available that express GFP fused to the selectable marker Zeocin™ . Each vector uses a different set of promoters to express the gene of interest and the Cycle 3 GFP-Zeocin™ fusion.

All three pTracer™ vectors have the following features:

• Cycle 3 GFP-Zeocin™ fusion for selection in mammalian cell lines
• BGH polyA signal and transcription termination sequences to enhance mRNA stability

The pTracer™-SV40 vector offers:
• SV40 promoter for high-level constitutive expression of your gene of interest
• CMV promoter for high-level constitutive expression of the Cycle 3 GFP-Zeocin™ fusion
• SV40 origin for episomal replication and simple vector rescue in cell lines expressing the SV40 large T antigen

The pTracer™-CMV2 vector offers:
• CMV promoter for high-level constitutive expression of your gene of interest
• EF-1α promoter for high-level constitutive expression of the Cycle 3 GFP-Zeocin™ fusion
• Ampicillin resistance gene for selection in E. coli

The pTracer™-EF vectors offer:
• EF-1α promoter for high-level constitutive expression of your gene of interest
• CMV promoter for high-level constitutive expression of the Cycle 3 GFP-Zeocin™ fusion
• C-terminal V5 epitope tag for simple detection of recombinant fusion proteins with an Anti-V5 Antibody
• C-terminal 6xHis tag for rapid purification on nickel-chelating resin and simple detection with an Anti-His(C-term) Antibody
• Multiple cloning site supplied in three reading frames to simplify cloning in frame with the C-terminal tag

pRSET A, B, & C Bacterial Expression Vectors (Invitrogen™)

The pRSET vector is designed for high-level prokaryotic expression controlled by the strong bacteriophage T7 promoter. Expression is induced by the production of T7 RNA polymerase in BL21(DE3) E. coli. These cells also produce T7 lysozyme to reduce basal expression of target genes. The pRSET vector offers:

• Bacteriophage T7 promoter for high-level expression
• T7 gene 10 sequence to provide protein stability
• N-terminal polyhistidine (6xHis) tag for rapid purification with nickel-chelating resin and detection with an Anti-HisG Antibody
• N-terminal Xpress™ epitope for detection with the Anti-Xpress™ Antibody
• Enterokinase cleavage site for removal of fusion tag

A set of three vectors is provided (A, B, and C). Each has the N-terminal tag coding sequence in a different reading frame relative to the multiple cloning site to simplify in-frame cloning of your gene.

pEF4/myc-His A, B, & C Mammalian Expression Vectors (Invitrogen™)

All pEF or pUB vectors contain a strong promoter for high-level expression in mammalian cells, a choice of selection marker for generating stable cell lines, and an epitope tag for easy detection with a monoclonal antibody and rapid purification on nickel-chelating resin. Each vector is available in three reading frames to simplify cloning in-frame with the fusion tag.

pcDNA™4/myc-His A, B, & C Mammalian Expression Vectors (Invitrogen™)

All pcDNA™ vectors contain a strong promoter for high-level expression in mammalian cells, a choice of selection marker for generating stable cell lines, and an epitope tag for easy detection with a monoclonal antibody and rapid purification on nickel-chelating resin. Each vector is available in three reading frames to simplify cloning in-frame with the fusion tag.

pMT/V5-His A, B, & C Drosophila Expression Vectors (Invitrogen™)

The DES®-Inducible Kit provides the expression vector pMT/V5-His for expression of recombinant proteins. This vector uses the Drosophila metallothionein gene promoter that is induced in S2 cells upon addition of copper sulfate or cadmium chloride to the culture medium.

The pMT/V5-His vector offers the following features:

• Small size (3.5 kb) to improve DNA yields and increase subcloning efficiency
• C-terminal V5 epitope tag for rapid detection with Invitrogen's Anti-V5 Antibody
• C-terminal 6xHis tag for simple purification of recombinant fusion proteins using nickel-chelating resin

To facilitate cloning, a set of three vectors-A, B, and C-is provided. Each vector has the multiple cloning site in a different reading frame relative to the coding sequence of the C-terminal tag.

pPIC6α A, B, & C Pichia Vectors (Invitrogen™)

pPIC6 A, B, & C are Pichia pastoris expression vectors designed for simple selection,
high-level expression, and rapid protein purification and detection. The pPIC6 A,
B, & C vectors are designed for high-level secreted expression. Both sets of vectors
have the following features:
Blasticidin resistance for direct selection of multi-copy integrants
Inducible AOX1 promoter for high-level expression in Pichia pastoris
C-terminal c-myc epitope for convenient detection with an Anti-myc Antibody
C-terminal polyhistidine (6xHis) tag for rapid purification with nickel-chelating
resin and detection with an Anti-His(C-term) Antibody
pPIC6 also features:
An É -factor secretion signal for efficient transport of proteins to the medium

pEF1/myc-His A, B, & C Mammalian Expression Vectors (Invitrogen™)

All pEF or pUB vectors contain a strong promoter for high-level expression in mammalian cells, a choice of selection marker for generating stable cell lines, and an epitope tag for easy detection with a monoclonal antibody and rapid purification on nickel-chelating resin. Each vector is available in three reading frames to simplify cloning in-frame with the fusion tag.

pSecTag2/Hygro A, B, & C Mammalian Expression Vectors (Invitrogen™)

pSecTag2 and pSecTag2/Hygro are mammalian expression vectors designed for the secretion, purification, and detection of fusion proteins. Each vector has a large multiple cloning site in three reading frames to simplify cloning in frame with the N-terminal secretion signal. The vectors (Figure 1) offer the following features:

• Secretion signal from the V-J2-C region of the mouse Ig kappa-chain for efficient secretion of recombinant proteins (Figure 2)
• Cytomegalovirus (CMV) promoter for high-level constitutive expression
• C-terminal polyhistidine (6xHis) tag for rapid purification with nickel-chelating resin and detection with an Anti-His(C-term) Antibody
• C-terminal c-myc epitope for detection with an Anti-myc Antibody
• Bovine Growth Hormone (BGH) polyadenylation signal and transcription termination sequence to enhance mRNA stability
• SV40 origin for episomal replication and simple vector rescue in cell lines expressing the large T antigen (e.g., COS-1, COS-7)

The pSecTag2 vectors carry the Zeocin™ resistance gene for cost-effective selection in mammalian cells. Zeocin™ selection can also be used in E. coli.

The pSecTag2/Hygro vectors have the Hygromycin B resistance gene for selection of stable mammalian cell lines.

pcDNA™3.1(-)/myc-His A, B, & C Mammalian Expression Vectors (Invitrogen™)

All pcDNA™ vectors contain a strong promoter for high-level expression in mammalian cells, a choice of selection marker for generating stable cell lines, and an epitope tag for easy detection with a monoclonal antibody and rapid purification on nickel-chelating resin. Each vector is available in three reading frames to simplify cloning in-frame with the fusion tag.

pGAPZ A, B, & C Pichia pastoris Expression Vectors (Invitrogen™)

pGAPZ A, B, & C and pGAPZ A, B, & C are expression vectors designed for high-level, constitutive expression in Pichia pastoris. pGAPZ and pGAPZ were created by replacing the methanol-regulated AOX1 promoter with the constitutive, glyceraldehyde-3-phosphate dehydrogenase (GAP) promoter in the backbone of the pPICZ vectors. Although the yield
of any protein constitutively expressed in the Pichia system is dependent on the toxicity of the protein to yeast, constitutive expression under the control of pGAPZ or pGAPZ vectors can sometimes produce greater yields than inducible expression (1).

pcDNA™4/V5-His A, B, & C Mammalian Expression Vectors (Invitrogen™)

All pcDNA™ vectors contain a strong promoter for high-level expression in mammalian cells, a choice of selection marker for generating stable cell lines, and an epitope tag for easy detection with a monoclonal antibody and rapid purification on nickel-chelating resin. Each vector is available in three reading frames to simplify cloning in-frame with the fusion tag.

pAc5.1/V5-His A, B, & C Vectors (Invitrogen™)

pAC5.1/V5-His vectors are designed for use in Drosophila cells to achieve high-level transient expression of recombinant proteins. The vector offers the strong, constitutive promoter from the Drosophila actin 5C gene. The pAC5.1/V5-His vectors can be used with the DES™-Inducible Kits (K512001 and K412001) for constitutive expression of your protein of interest. The pAc5.1/V5-His vectors also offer the following features:

• Small size (5.4 kb) to improve DNA yields and increase subcloning efficiency
• C-terminal V5 epitope tag for rapid detection with Invitrogens Anti-V5 Antibody
• C-terminal 6xHis tag for simple purification of recombinant fusion proteins using nickel-chelating resin

To facilitate cloning, a set of three vectors-A, B, and C-is provided. Each vector has the multiple cloning site in a different reading frame relative to the coding sequence of the C-terminal tag.

pcDNA™3.1/V5-His A, B, & C Mammalian Expression Vectors (Invitrogen™)

All pcDNA™ vectors contain a strong promoter for high-level expression in mammalian cells, a choice of selection marker for generating stable cell lines, and an epitope tag for easy detection with a monoclonal antibody and rapid purification on nickel-chelating resin. Each vector is available in three reading frames to simplify cloning in-frame with the fusion tag.