atg deficient frt hygro fusion gene  (Thermo Fisher)


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    Name:
    pSecTag2 Hygro A B C Mammalian Expression Vectors
    Description:
    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
    Catalog Number:
    v91020
    Price:
    None
    Category:
    DNA Vectors Clones Purified Nucleic Acids Libraries
    Applications:
    Constitutive Expression|Mammalian Expression|Protein Biology|Protein Expression|Secreted Expression
    Buy from Supplier


    Structured Review

    Thermo Fisher atg deficient frt hygro fusion gene
    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
    https://www.bioz.com/result/atg deficient frt hygro fusion gene/product/Thermo Fisher
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    atg deficient frt hygro fusion gene - by Bioz Stars, 2021-03
    97/100 stars

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    Related Articles

    Construct:

    Article Title: A peptide derived from an extracellular domain selectively inhibits receptor internalization: Target sequences on insulin and insulin-like growth factor 1 receptors
    Article Snippet: IRp binding to IGF-1R was carried out in the presence or absence of 5 nM insulin or IGF-1. .. Deletion mutants were constructed using the PCR and standard cloning techniques , and subcloned into a mammalian expression vector pCR3.1 (Invitrogen). .. A 2.0-kb fragment of the C-terminal portion of the gene (starting at L696 ) was created by using the PCR primers: IRC, 5′-CCCG T C TAGA TAGGCACTGTTAGGAAGG-3′ and IRMP2, 5′- A TGGAGGAGT A CTCGTTT AAA AAGACGTTTG G G CCC TACCTGCACAACGT TAA CTTCGTCCCCAGGCCATCTCGG-3′.

    Polymerase Chain Reaction:

    Article Title: A peptide derived from an extracellular domain selectively inhibits receptor internalization: Target sequences on insulin and insulin-like growth factor 1 receptors
    Article Snippet: IRp binding to IGF-1R was carried out in the presence or absence of 5 nM insulin or IGF-1. .. Deletion mutants were constructed using the PCR and standard cloning techniques , and subcloned into a mammalian expression vector pCR3.1 (Invitrogen). .. A 2.0-kb fragment of the C-terminal portion of the gene (starting at L696 ) was created by using the PCR primers: IRC, 5′-CCCG T C TAGA TAGGCACTGTTAGGAAGG-3′ and IRMP2, 5′- A TGGAGGAGT A CTCGTTT AAA AAGACGTTTG G G CCC TACCTGCACAACGT TAA CTTCGTCCCCAGGCCATCTCGG-3′.

    Article Title: Technical Brief: Subretinal injection and electroporation into adult mouse eyes
    Article Snippet: .. Reporter gene plasmid The base vector was the mammalian expression vector pVAX™200-DEST (Invitrogen, Carlsbad, CA). tdTomato cDNA was PCR amplified from pRSETB-tdTomato with primers bearing AttB sites and incubated with BP ClonaseII and the pVAX™200-DEST plasmid. .. The resulting reporter expression plasmid, called pVAX-tdTomato [ ], contained the CMV Immediate Early promoter driving expression of tdTomato.

    Clone Assay:

    Article Title: A peptide derived from an extracellular domain selectively inhibits receptor internalization: Target sequences on insulin and insulin-like growth factor 1 receptors
    Article Snippet: IRp binding to IGF-1R was carried out in the presence or absence of 5 nM insulin or IGF-1. .. Deletion mutants were constructed using the PCR and standard cloning techniques , and subcloned into a mammalian expression vector pCR3.1 (Invitrogen). .. A 2.0-kb fragment of the C-terminal portion of the gene (starting at L696 ) was created by using the PCR primers: IRC, 5′-CCCG T C TAGA TAGGCACTGTTAGGAAGG-3′ and IRMP2, 5′- A TGGAGGAGT A CTCGTTT AAA AAGACGTTTG G G CCC TACCTGCACAACGT TAA CTTCGTCCCCAGGCCATCTCGG-3′.

    Article Title: Pathogenic Natural Antibodies Recognizing Annexin IV Are Required to Develop Intestinal Ischemia-Reperfusion Injury
    Article Snippet: The KpnI/XhoI fragment was amplified by PCR from genomic DNA using Pfu polymerase (Novagen). .. The KpnI/XhoI fragment was subsequently cloned into pSecTag2/Hygro B expression vector (Invitrogen) and expression of the protein was accomplished in the F-293 cell line (Invitrogen). .. The vector adds an Ig k-chain leader sequence to the N-terminus of the protein for secretion.

    Article Title: SDF-1? degrades, while gp120 upregulates BimEL: implications for the development of T cell memory
    Article Snippet: The N-terminus BimS construct was PCR amplified using a sense primer with BamH1 site (5′- CGGATCC ATGGCAAAGCAA) coupled with antisense splicing primer (5′-GGAAGCTTGTGGCTC) and C-terminus BimS was PCR amplified using a antisense primer with EcoR1 site (5′- CGAATTC TCAATGCATTCT) coupled with sense splicing primer (5′-CCACAAGCTTCCATGAGG). .. The BimEL , BimL and BimS were cloned into either mammalian expression vector pEF1 (Invitrogen) to express full size Bim proteins or into GFP-pCDNA3 (Invitrogen) to express GFP-tagged Bim proteins. .. All Bim constructs were confirmed by DNA sequence analysis and tested for expression prior to experimental use.

    Expressing:

    Article Title: A peptide derived from an extracellular domain selectively inhibits receptor internalization: Target sequences on insulin and insulin-like growth factor 1 receptors
    Article Snippet: IRp binding to IGF-1R was carried out in the presence or absence of 5 nM insulin or IGF-1. .. Deletion mutants were constructed using the PCR and standard cloning techniques , and subcloned into a mammalian expression vector pCR3.1 (Invitrogen). .. A 2.0-kb fragment of the C-terminal portion of the gene (starting at L696 ) was created by using the PCR primers: IRC, 5′-CCCG T C TAGA TAGGCACTGTTAGGAAGG-3′ and IRMP2, 5′- A TGGAGGAGT A CTCGTTT AAA AAGACGTTTG G G CCC TACCTGCACAACGT TAA CTTCGTCCCCAGGCCATCTCGG-3′.

    Article Title: The functional expression of toxic genes: Lessons learned from molecular cloning of CCH1, a high-affinity Ca2+ channel
    Article Snippet: HEK293 cells were purchased from ATCC (CRL-1573). .. The aim was to clone CCH1 into the pcDNA3.1/CT-GFP-topo expression plasmid under the control of the constitutive mammalian CMV promoter (Invitrogen, Carlsbad CA). .. To circumvent propagating the expression plasmid in E. coli , the essential features of pcDNA3.1/CT-GFP-topo that were needed for the expression of CCH1 in HEK293 cells were PCR-amplified and cloned into a yeast expression plasmid, pRS425.

    Article Title: Pathogenic Natural Antibodies Recognizing Annexin IV Are Required to Develop Intestinal Ischemia-Reperfusion Injury
    Article Snippet: The KpnI/XhoI fragment was amplified by PCR from genomic DNA using Pfu polymerase (Novagen). .. The KpnI/XhoI fragment was subsequently cloned into pSecTag2/Hygro B expression vector (Invitrogen) and expression of the protein was accomplished in the F-293 cell line (Invitrogen). .. The vector adds an Ig k-chain leader sequence to the N-terminus of the protein for secretion.

    Article Title: Mononuclear Cell-Infiltrate Inhibition by Blocking Macrophage-Derived Chemokine Results in Attenuation of Developing Crescentic Glomerulonephritis
    Article Snippet: A polyclonal antiserum was raised by immunizing a rabbit with recombinant MDC or CCR4 following previously described procedures. .. The full-length rat MDC and CCR4 were subcloned into the mammalian cell expression vector pCDM8 (Invitrogen) and transfected into subconfluent 293T cells by electroporation. .. After 48 hours of incubation, the supernatant or the cell pellet were collected, electrophoresed in a 4 to 12% Bis-Tris Gel with 2-(N-morpholino) ethane sulfonic acid (MES) as a running buffer, transferred to nitrocellulose membrane, and blotted with various antisera.

    Article Title: c-Myc Transformation Domain Recruits the Human STAGA Complex and Requires TRRAP and GCN5 Acetylase Activity for Transcription Activation*
    Article Snippet: HEK293 cells were seeded on 10-cm plates in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum (without antibiotics) at a density of 6 × 106 24 h before transfection. .. Cells at ~90% confluence were transfected with 10 μ g of CMV expression vectors encoding FLAG-tagged mouse Myc full-length or Δ1–110 using LipofectAMINE 2000 reagent (Invitrogen). ..

    Article Title: SDF-1? degrades, while gp120 upregulates BimEL: implications for the development of T cell memory
    Article Snippet: The N-terminus BimS construct was PCR amplified using a sense primer with BamH1 site (5′- CGGATCC ATGGCAAAGCAA) coupled with antisense splicing primer (5′-GGAAGCTTGTGGCTC) and C-terminus BimS was PCR amplified using a antisense primer with EcoR1 site (5′- CGAATTC TCAATGCATTCT) coupled with sense splicing primer (5′-CCACAAGCTTCCATGAGG). .. The BimEL , BimL and BimS were cloned into either mammalian expression vector pEF1 (Invitrogen) to express full size Bim proteins or into GFP-pCDNA3 (Invitrogen) to express GFP-tagged Bim proteins. .. All Bim constructs were confirmed by DNA sequence analysis and tested for expression prior to experimental use.

    Article Title: Technical Brief: Subretinal injection and electroporation into adult mouse eyes
    Article Snippet: .. Reporter gene plasmid The base vector was the mammalian expression vector pVAX™200-DEST (Invitrogen, Carlsbad, CA). tdTomato cDNA was PCR amplified from pRSETB-tdTomato with primers bearing AttB sites and incubated with BP ClonaseII and the pVAX™200-DEST plasmid. .. The resulting reporter expression plasmid, called pVAX-tdTomato [ ], contained the CMV Immediate Early promoter driving expression of tdTomato.

    Article Title: The human cytomegalovirus protein UL116 interacts with the viral ER resident glycoprotein UL148 and promotes the incorporation of gH/gL complexes into virions
    Article Snippet: The next day, the Protein G magnetic beads were washed three times in lysis buffer, and eluted by heating (50°C, 10 min) in 2×Laemmli buffer (4% SDS, 20% glycerol, 0.004% bromphenol blue, 0.125 M Tris HCl, pH 6.8) lacking reducing agent. .. To obtain starting material for IP experiments from transfected cells, 3 μg of mammalian expression vector plasmids based on pEF1α V5 His C (Invitrogen) encoding codon-optimized gH, UL116myc , or UL148HA were transfected using 9 μL of TransIT-2020 reagent (Mirus, Inc, #MIR 5404) into 6 × 105 HEK-293T cells that had been seeded into wells of a six-well cluster plate. .. As needed, transfections were supplemented with empty pEF1α V5 His C vector to maintain equivalent amounts of DNA per transfection reaction.

    Plasmid Preparation:

    Article Title: A peptide derived from an extracellular domain selectively inhibits receptor internalization: Target sequences on insulin and insulin-like growth factor 1 receptors
    Article Snippet: IRp binding to IGF-1R was carried out in the presence or absence of 5 nM insulin or IGF-1. .. Deletion mutants were constructed using the PCR and standard cloning techniques , and subcloned into a mammalian expression vector pCR3.1 (Invitrogen). .. A 2.0-kb fragment of the C-terminal portion of the gene (starting at L696 ) was created by using the PCR primers: IRC, 5′-CCCG T C TAGA TAGGCACTGTTAGGAAGG-3′ and IRMP2, 5′- A TGGAGGAGT A CTCGTTT AAA AAGACGTTTG G G CCC TACCTGCACAACGT TAA CTTCGTCCCCAGGCCATCTCGG-3′.

    Article Title: The functional expression of toxic genes: Lessons learned from molecular cloning of CCH1, a high-affinity Ca2+ channel
    Article Snippet: HEK293 cells were purchased from ATCC (CRL-1573). .. The aim was to clone CCH1 into the pcDNA3.1/CT-GFP-topo expression plasmid under the control of the constitutive mammalian CMV promoter (Invitrogen, Carlsbad CA). .. To circumvent propagating the expression plasmid in E. coli , the essential features of pcDNA3.1/CT-GFP-topo that were needed for the expression of CCH1 in HEK293 cells were PCR-amplified and cloned into a yeast expression plasmid, pRS425.

    Article Title: Pathogenic Natural Antibodies Recognizing Annexin IV Are Required to Develop Intestinal Ischemia-Reperfusion Injury
    Article Snippet: The KpnI/XhoI fragment was amplified by PCR from genomic DNA using Pfu polymerase (Novagen). .. The KpnI/XhoI fragment was subsequently cloned into pSecTag2/Hygro B expression vector (Invitrogen) and expression of the protein was accomplished in the F-293 cell line (Invitrogen). .. The vector adds an Ig k-chain leader sequence to the N-terminus of the protein for secretion.

    Article Title: Mononuclear Cell-Infiltrate Inhibition by Blocking Macrophage-Derived Chemokine Results in Attenuation of Developing Crescentic Glomerulonephritis
    Article Snippet: A polyclonal antiserum was raised by immunizing a rabbit with recombinant MDC or CCR4 following previously described procedures. .. The full-length rat MDC and CCR4 were subcloned into the mammalian cell expression vector pCDM8 (Invitrogen) and transfected into subconfluent 293T cells by electroporation. .. After 48 hours of incubation, the supernatant or the cell pellet were collected, electrophoresed in a 4 to 12% Bis-Tris Gel with 2-(N-morpholino) ethane sulfonic acid (MES) as a running buffer, transferred to nitrocellulose membrane, and blotted with various antisera.

    Article Title: SDF-1? degrades, while gp120 upregulates BimEL: implications for the development of T cell memory
    Article Snippet: The N-terminus BimS construct was PCR amplified using a sense primer with BamH1 site (5′- CGGATCC ATGGCAAAGCAA) coupled with antisense splicing primer (5′-GGAAGCTTGTGGCTC) and C-terminus BimS was PCR amplified using a antisense primer with EcoR1 site (5′- CGAATTC TCAATGCATTCT) coupled with sense splicing primer (5′-CCACAAGCTTCCATGAGG). .. The BimEL , BimL and BimS were cloned into either mammalian expression vector pEF1 (Invitrogen) to express full size Bim proteins or into GFP-pCDNA3 (Invitrogen) to express GFP-tagged Bim proteins. .. All Bim constructs were confirmed by DNA sequence analysis and tested for expression prior to experimental use.

    Article Title: Technical Brief: Subretinal injection and electroporation into adult mouse eyes
    Article Snippet: .. Reporter gene plasmid The base vector was the mammalian expression vector pVAX™200-DEST (Invitrogen, Carlsbad, CA). tdTomato cDNA was PCR amplified from pRSETB-tdTomato with primers bearing AttB sites and incubated with BP ClonaseII and the pVAX™200-DEST plasmid. .. The resulting reporter expression plasmid, called pVAX-tdTomato [ ], contained the CMV Immediate Early promoter driving expression of tdTomato.

    Article Title: The human cytomegalovirus protein UL116 interacts with the viral ER resident glycoprotein UL148 and promotes the incorporation of gH/gL complexes into virions
    Article Snippet: The next day, the Protein G magnetic beads were washed three times in lysis buffer, and eluted by heating (50°C, 10 min) in 2×Laemmli buffer (4% SDS, 20% glycerol, 0.004% bromphenol blue, 0.125 M Tris HCl, pH 6.8) lacking reducing agent. .. To obtain starting material for IP experiments from transfected cells, 3 μg of mammalian expression vector plasmids based on pEF1α V5 His C (Invitrogen) encoding codon-optimized gH, UL116myc , or UL148HA were transfected using 9 μL of TransIT-2020 reagent (Mirus, Inc, #MIR 5404) into 6 × 105 HEK-293T cells that had been seeded into wells of a six-well cluster plate. .. As needed, transfections were supplemented with empty pEF1α V5 His C vector to maintain equivalent amounts of DNA per transfection reaction.

    Transfection:

    Article Title: Mononuclear Cell-Infiltrate Inhibition by Blocking Macrophage-Derived Chemokine Results in Attenuation of Developing Crescentic Glomerulonephritis
    Article Snippet: A polyclonal antiserum was raised by immunizing a rabbit with recombinant MDC or CCR4 following previously described procedures. .. The full-length rat MDC and CCR4 were subcloned into the mammalian cell expression vector pCDM8 (Invitrogen) and transfected into subconfluent 293T cells by electroporation. .. After 48 hours of incubation, the supernatant or the cell pellet were collected, electrophoresed in a 4 to 12% Bis-Tris Gel with 2-(N-morpholino) ethane sulfonic acid (MES) as a running buffer, transferred to nitrocellulose membrane, and blotted with various antisera.

    Article Title: c-Myc Transformation Domain Recruits the Human STAGA Complex and Requires TRRAP and GCN5 Acetylase Activity for Transcription Activation*
    Article Snippet: HEK293 cells were seeded on 10-cm plates in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum (without antibiotics) at a density of 6 × 106 24 h before transfection. .. Cells at ~90% confluence were transfected with 10 μ g of CMV expression vectors encoding FLAG-tagged mouse Myc full-length or Δ1–110 using LipofectAMINE 2000 reagent (Invitrogen). ..

    Article Title: The human cytomegalovirus protein UL116 interacts with the viral ER resident glycoprotein UL148 and promotes the incorporation of gH/gL complexes into virions
    Article Snippet: The next day, the Protein G magnetic beads were washed three times in lysis buffer, and eluted by heating (50°C, 10 min) in 2×Laemmli buffer (4% SDS, 20% glycerol, 0.004% bromphenol blue, 0.125 M Tris HCl, pH 6.8) lacking reducing agent. .. To obtain starting material for IP experiments from transfected cells, 3 μg of mammalian expression vector plasmids based on pEF1α V5 His C (Invitrogen) encoding codon-optimized gH, UL116myc , or UL148HA were transfected using 9 μL of TransIT-2020 reagent (Mirus, Inc, #MIR 5404) into 6 × 105 HEK-293T cells that had been seeded into wells of a six-well cluster plate. .. As needed, transfections were supplemented with empty pEF1α V5 His C vector to maintain equivalent amounts of DNA per transfection reaction.

    Electroporation:

    Article Title: Mononuclear Cell-Infiltrate Inhibition by Blocking Macrophage-Derived Chemokine Results in Attenuation of Developing Crescentic Glomerulonephritis
    Article Snippet: A polyclonal antiserum was raised by immunizing a rabbit with recombinant MDC or CCR4 following previously described procedures. .. The full-length rat MDC and CCR4 were subcloned into the mammalian cell expression vector pCDM8 (Invitrogen) and transfected into subconfluent 293T cells by electroporation. .. After 48 hours of incubation, the supernatant or the cell pellet were collected, electrophoresed in a 4 to 12% Bis-Tris Gel with 2-(N-morpholino) ethane sulfonic acid (MES) as a running buffer, transferred to nitrocellulose membrane, and blotted with various antisera.

    Amplification:

    Article Title: Technical Brief: Subretinal injection and electroporation into adult mouse eyes
    Article Snippet: .. Reporter gene plasmid The base vector was the mammalian expression vector pVAX™200-DEST (Invitrogen, Carlsbad, CA). tdTomato cDNA was PCR amplified from pRSETB-tdTomato with primers bearing AttB sites and incubated with BP ClonaseII and the pVAX™200-DEST plasmid. .. The resulting reporter expression plasmid, called pVAX-tdTomato [ ], contained the CMV Immediate Early promoter driving expression of tdTomato.

    Incubation:

    Article Title: Technical Brief: Subretinal injection and electroporation into adult mouse eyes
    Article Snippet: .. Reporter gene plasmid The base vector was the mammalian expression vector pVAX™200-DEST (Invitrogen, Carlsbad, CA). tdTomato cDNA was PCR amplified from pRSETB-tdTomato with primers bearing AttB sites and incubated with BP ClonaseII and the pVAX™200-DEST plasmid. .. The resulting reporter expression plasmid, called pVAX-tdTomato [ ], contained the CMV Immediate Early promoter driving expression of tdTomato.

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  • 97
    Thermo Fisher atg deficient frt hygro fusion gene
    Atg Deficient Frt Hygro Fusion Gene, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/atg deficient frt hygro fusion gene/product/Thermo Fisher
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    atg deficient frt hygro fusion gene - by Bioz Stars, 2021-03
    97/100 stars
      Buy from Supplier

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