pcdna dest47  (Thermo Fisher)


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    Name:
    Gateway pcDNA DEST47 Vector
    Description:
    The pcDNA vectors are designed for high level constitutive expression in a variety of mammalian cell lines The Gateway pcDNA DEST47 vector offers the following key features •C terminal Cycle 3 GFP tag for rapid detection of recombinant protein•Cytomegalovirus CMV promoter for high level expression•attR sites for Gateway cloning enabling recombination with attL flanked fragments•Neomycin resistance gene for stable selection•Ampicillin resistance gene and pUC origin for selection and maintenance in E coliGateway CloningTo 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 ORF Clone The following table lists a variety of available destination vectors Additional materials required available separately Gateway entry clone appropriate Gateway LR Clonase enzyme mix and reaction buffer
    Catalog Number:
    12281010
    Price:
    None
    Applications:
    Cloning|Constitutive Expression|Destination Vectors|Gateway Cloning|Mammalian Expression|Protein Biology|Protein Expression
    Category:
    DNA Vectors Clones Purified Nucleic Acids Libraries
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    Structured Review

    Thermo Fisher pcdna dest47
    The pcDNA vectors are designed for high level constitutive expression in a variety of mammalian cell lines The Gateway pcDNA DEST47 vector offers the following key features •C terminal Cycle 3 GFP tag for rapid detection of recombinant protein•Cytomegalovirus CMV promoter for high level expression•attR sites for Gateway cloning enabling recombination with attL flanked fragments•Neomycin resistance gene for stable selection•Ampicillin resistance gene and pUC origin for selection and maintenance in E coliGateway CloningTo 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 ORF Clone The following table lists a variety of available destination vectors Additional materials required available separately Gateway entry clone appropriate Gateway LR Clonase enzyme mix and reaction buffer
    https://www.bioz.com/result/pcdna dest47/product/Thermo Fisher
    Average 94 stars, based on 21 article reviews
    Price from $9.99 to $1999.99
    pcdna dest47 - by Bioz Stars, 2020-09
    94/100 stars

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    Related Articles

    Clone Assay:

    Article Title: De novo phosphorylation of H2AX by WSTF regulates transcription-coupled homologous recombination repair
    Article Snippet: .. Then the clones into pENTRY were sequenced and transferred pcDNA-DEST47 (GFP C-terminal; Invitrogen), pcDNA-DEST53 (GFP N-terminal; Invitrogen) or pcDNA-DEST-FLAG vector using a Gateway LR Cloning system (Invitrogen). .. To generate WSTF and histone H2AX mutant plasmids, site-directed mutagenesis were carried out using the QuickChange site-directed mutagenesis kit (Stratagene) or by a classical PCR method using pENTRY clones.

    Article Title: Deficiency of the Cytoskeletal Protein SPECC1L Leads to Oblique Facial Clefting
    Article Snippet: .. Full-length and C-terminal calponin homology domain (CHD) truncated cDNA (designated ΔCHD) of Specc1l ( ) were cloned into a Gateway pENTR/SD/D-TOPO vector (Invitrogen, Carlsbad, CA) and transferred to a Gateway-compatible destination vector with a C-terminal GFP tag (pcDNA-DEST47, Invitrogen, Carlsbad, CA). p.Thr190Met and p.Gln415Pro mutations were created with the QuikChange Mutagenesis kit (Stratagene, La Jolla, CA) according to the manufacturer's protocol. .. The cDNA fragment encoding amino acids 887–997 of SPECC1L was cloned into the pGEX6p-2 vector to create a GST-tagged peptide that was then used to generate a rabbit polyclonal antibody (Covance, Princeton, NJ).

    Article Title: Ca2+ Conduction by an Amino Acid-Gated Ion Channel Related to Glutamate Receptors 1 Conduction by an Amino Acid-Gated Ion Channel Related to Glutamate Receptors 1 [W]
    Article Snippet: .. The resulting PCR fragment was cloned into the pENTR- d entry vector (Invitrogen) followed by a GATEWAY recombination reaction (Invitrogen) into pcDNA-DEST47 mammalian expression vector (pcDEST47-GLR3.4-GFP; for transfection of 293T cells) or the modified pEARLEYGATE 102 plant expression vector ( ) in which the cyan fluorescent protein fragment was replaced with EGFP for enhanced fluorescence to produce the respective 35S:GLR3.4-EGFP . .. The floral dip method was used to transform the glr3.4-1 mutant with the 35S:GLR3.4-EGFP construct.

    Article Title: The Ciliary Protein Arl13b Functions Outside of the Primary Cilium in Shh-Mediated Axon Guidance
    Article Snippet: .. The shRNA/miR/EmGFP sequence was then cloned via attB/attP site recombination (using BP clonase) into the pDONR221 backbone (ThermoFisher, 12536-017), and via attL/attR site recombination (using LR clonase) into a custom expression vector derived from Gateway pcDNA-DEST47 (ThermoFisher Scientific, 12281-010). .. This custom expression vector replaced the promoter and polyA regions of Gateway pcDNA-DEST47 with the promoter and polyA from the pCAGGS vector ( ) to improve its expression in mammalian neurons.

    Transfection:

    Article Title: Ca2+ Conduction by an Amino Acid-Gated Ion Channel Related to Glutamate Receptors 1 Conduction by an Amino Acid-Gated Ion Channel Related to Glutamate Receptors 1 [W]
    Article Snippet: .. The resulting PCR fragment was cloned into the pENTR- d entry vector (Invitrogen) followed by a GATEWAY recombination reaction (Invitrogen) into pcDNA-DEST47 mammalian expression vector (pcDEST47-GLR3.4-GFP; for transfection of 293T cells) or the modified pEARLEYGATE 102 plant expression vector ( ) in which the cyan fluorescent protein fragment was replaced with EGFP for enhanced fluorescence to produce the respective 35S:GLR3.4-EGFP . .. The floral dip method was used to transform the glr3.4-1 mutant with the 35S:GLR3.4-EGFP construct.

    shRNA:

    Article Title: The Ciliary Protein Arl13b Functions Outside of the Primary Cilium in Shh-Mediated Axon Guidance
    Article Snippet: .. The shRNA/miR/EmGFP sequence was then cloned via attB/attP site recombination (using BP clonase) into the pDONR221 backbone (ThermoFisher, 12536-017), and via attL/attR site recombination (using LR clonase) into a custom expression vector derived from Gateway pcDNA-DEST47 (ThermoFisher Scientific, 12281-010). .. This custom expression vector replaced the promoter and polyA regions of Gateway pcDNA-DEST47 with the promoter and polyA from the pCAGGS vector ( ) to improve its expression in mammalian neurons.

    Fluorescence:

    Article Title: Ca2+ Conduction by an Amino Acid-Gated Ion Channel Related to Glutamate Receptors 1 Conduction by an Amino Acid-Gated Ion Channel Related to Glutamate Receptors 1 [W]
    Article Snippet: .. The resulting PCR fragment was cloned into the pENTR- d entry vector (Invitrogen) followed by a GATEWAY recombination reaction (Invitrogen) into pcDNA-DEST47 mammalian expression vector (pcDEST47-GLR3.4-GFP; for transfection of 293T cells) or the modified pEARLEYGATE 102 plant expression vector ( ) in which the cyan fluorescent protein fragment was replaced with EGFP for enhanced fluorescence to produce the respective 35S:GLR3.4-EGFP . .. The floral dip method was used to transform the glr3.4-1 mutant with the 35S:GLR3.4-EGFP construct.

    Polymerase Chain Reaction:

    Article Title: Rapid Aquaporin Translocation Regulates Cellular Water Flow
    Article Snippet: .. Samples were heated to 94 °C for 2 min, followed by 30 cycles of 94 °C for 30 s, 55 °C for 30 s, and 68 °C for 3 min, and then 68 °C for 7 min. Purified PCR products were subcloned into the pDONR221TM entry vector (Invitrogen) using the att B1 and att B2 sites in a reaction with GatewayTM BP ClonaseTM enzyme mix (Invitrogen). pDONR221TM vectors containing the required sequences were recombined with the pcDNA-DEST47 GatewayTM vector using the att L and att R reaction with GatewayTM LR ClonaseTM enzyme mix (Invitrogen). .. This created expression vectors with the cycle 3 mutant of the GFP gene at the carboxyl terminus of the AQP gene of interest, which were expressed subsequently as fusion proteins.

    Article Title: Ca2+ Conduction by an Amino Acid-Gated Ion Channel Related to Glutamate Receptors 1 Conduction by an Amino Acid-Gated Ion Channel Related to Glutamate Receptors 1 [W]
    Article Snippet: .. The resulting PCR fragment was cloned into the pENTR- d entry vector (Invitrogen) followed by a GATEWAY recombination reaction (Invitrogen) into pcDNA-DEST47 mammalian expression vector (pcDEST47-GLR3.4-GFP; for transfection of 293T cells) or the modified pEARLEYGATE 102 plant expression vector ( ) in which the cyan fluorescent protein fragment was replaced with EGFP for enhanced fluorescence to produce the respective 35S:GLR3.4-EGFP . .. The floral dip method was used to transform the glr3.4-1 mutant with the 35S:GLR3.4-EGFP construct.

    Mutagenesis:

    Article Title: Deficiency of the Cytoskeletal Protein SPECC1L Leads to Oblique Facial Clefting
    Article Snippet: .. Full-length and C-terminal calponin homology domain (CHD) truncated cDNA (designated ΔCHD) of Specc1l ( ) were cloned into a Gateway pENTR/SD/D-TOPO vector (Invitrogen, Carlsbad, CA) and transferred to a Gateway-compatible destination vector with a C-terminal GFP tag (pcDNA-DEST47, Invitrogen, Carlsbad, CA). p.Thr190Met and p.Gln415Pro mutations were created with the QuikChange Mutagenesis kit (Stratagene, La Jolla, CA) according to the manufacturer's protocol. .. The cDNA fragment encoding amino acids 887–997 of SPECC1L was cloned into the pGEX6p-2 vector to create a GST-tagged peptide that was then used to generate a rabbit polyclonal antibody (Covance, Princeton, NJ).

    Purification:

    Article Title: Rapid Aquaporin Translocation Regulates Cellular Water Flow
    Article Snippet: .. Samples were heated to 94 °C for 2 min, followed by 30 cycles of 94 °C for 30 s, 55 °C for 30 s, and 68 °C for 3 min, and then 68 °C for 7 min. Purified PCR products were subcloned into the pDONR221TM entry vector (Invitrogen) using the att B1 and att B2 sites in a reaction with GatewayTM BP ClonaseTM enzyme mix (Invitrogen). pDONR221TM vectors containing the required sequences were recombined with the pcDNA-DEST47 GatewayTM vector using the att L and att R reaction with GatewayTM LR ClonaseTM enzyme mix (Invitrogen). .. This created expression vectors with the cycle 3 mutant of the GFP gene at the carboxyl terminus of the AQP gene of interest, which were expressed subsequently as fusion proteins.

    Article Title: Concatenation of Human Connexin26 (hCx26) and Human Connexin46 (hCx46) for the Analysis of Heteromeric Gap Junction Hemichannels and Heterotypic Gap Junction Channels
    Article Snippet: .. The purified psDEST47, which is similar to the commercially available pcDNA™-DEST47 vector (#12281010, Thermo Fisher Scientific, Waltham, MA, USA), was used to create a C-terminally GFP-labeled fusion protein via the LR-cloning reaction. .. The vector psGEMHE-GW was used for in vitro transcription to produce the cRNA for the Xenopus oocytes.

    Modification:

    Article Title: Ca2+ Conduction by an Amino Acid-Gated Ion Channel Related to Glutamate Receptors 1 Conduction by an Amino Acid-Gated Ion Channel Related to Glutamate Receptors 1 [W]
    Article Snippet: .. The resulting PCR fragment was cloned into the pENTR- d entry vector (Invitrogen) followed by a GATEWAY recombination reaction (Invitrogen) into pcDNA-DEST47 mammalian expression vector (pcDEST47-GLR3.4-GFP; for transfection of 293T cells) or the modified pEARLEYGATE 102 plant expression vector ( ) in which the cyan fluorescent protein fragment was replaced with EGFP for enhanced fluorescence to produce the respective 35S:GLR3.4-EGFP . .. The floral dip method was used to transform the glr3.4-1 mutant with the 35S:GLR3.4-EGFP construct.

    Plasmid Preparation:

    Article Title: De novo phosphorylation of H2AX by WSTF regulates transcription-coupled homologous recombination repair
    Article Snippet: .. Then the clones into pENTRY were sequenced and transferred pcDNA-DEST47 (GFP C-terminal; Invitrogen), pcDNA-DEST53 (GFP N-terminal; Invitrogen) or pcDNA-DEST-FLAG vector using a Gateway LR Cloning system (Invitrogen). .. To generate WSTF and histone H2AX mutant plasmids, site-directed mutagenesis were carried out using the QuickChange site-directed mutagenesis kit (Stratagene) or by a classical PCR method using pENTRY clones.

    Article Title: Deficiency of the Cytoskeletal Protein SPECC1L Leads to Oblique Facial Clefting
    Article Snippet: .. Full-length and C-terminal calponin homology domain (CHD) truncated cDNA (designated ΔCHD) of Specc1l ( ) were cloned into a Gateway pENTR/SD/D-TOPO vector (Invitrogen, Carlsbad, CA) and transferred to a Gateway-compatible destination vector with a C-terminal GFP tag (pcDNA-DEST47, Invitrogen, Carlsbad, CA). p.Thr190Met and p.Gln415Pro mutations were created with the QuikChange Mutagenesis kit (Stratagene, La Jolla, CA) according to the manufacturer's protocol. .. The cDNA fragment encoding amino acids 887–997 of SPECC1L was cloned into the pGEX6p-2 vector to create a GST-tagged peptide that was then used to generate a rabbit polyclonal antibody (Covance, Princeton, NJ).

    Article Title: Enigma proteins regulate YAP mechanotransduction
    Article Snippet: .. To constitutively overexpress human PDLIM7, the PDLIM7 (isoform 1) open-reading frame (ORF) was subcloned from the corresponding entry vector (Dharmacon; clone 3562 for PDLIM7 isoform 1) into the destination vector pcDNA-PDEST47 (Invitrogen, 12281010) by recombination using the Gateway LR clonase enzyme mix (Invitrogen,11791). .. Human cell culture Human Caco-2 cells and HEK293T (Francis Crick Institute cell services) were grown in conditions as previously described ( ).

    Article Title: Rapid Aquaporin Translocation Regulates Cellular Water Flow
    Article Snippet: .. Samples were heated to 94 °C for 2 min, followed by 30 cycles of 94 °C for 30 s, 55 °C for 30 s, and 68 °C for 3 min, and then 68 °C for 7 min. Purified PCR products were subcloned into the pDONR221TM entry vector (Invitrogen) using the att B1 and att B2 sites in a reaction with GatewayTM BP ClonaseTM enzyme mix (Invitrogen). pDONR221TM vectors containing the required sequences were recombined with the pcDNA-DEST47 GatewayTM vector using the att L and att R reaction with GatewayTM LR ClonaseTM enzyme mix (Invitrogen). .. This created expression vectors with the cycle 3 mutant of the GFP gene at the carboxyl terminus of the AQP gene of interest, which were expressed subsequently as fusion proteins.

    Article Title: Ca2+ Conduction by an Amino Acid-Gated Ion Channel Related to Glutamate Receptors 1 Conduction by an Amino Acid-Gated Ion Channel Related to Glutamate Receptors 1 [W]
    Article Snippet: .. The resulting PCR fragment was cloned into the pENTR- d entry vector (Invitrogen) followed by a GATEWAY recombination reaction (Invitrogen) into pcDNA-DEST47 mammalian expression vector (pcDEST47-GLR3.4-GFP; for transfection of 293T cells) or the modified pEARLEYGATE 102 plant expression vector ( ) in which the cyan fluorescent protein fragment was replaced with EGFP for enhanced fluorescence to produce the respective 35S:GLR3.4-EGFP . .. The floral dip method was used to transform the glr3.4-1 mutant with the 35S:GLR3.4-EGFP construct.

    Article Title: The Ciliary Protein Arl13b Functions Outside of the Primary Cilium in Shh-Mediated Axon Guidance
    Article Snippet: .. The shRNA/miR/EmGFP sequence was then cloned via attB/attP site recombination (using BP clonase) into the pDONR221 backbone (ThermoFisher, 12536-017), and via attL/attR site recombination (using LR clonase) into a custom expression vector derived from Gateway pcDNA-DEST47 (ThermoFisher Scientific, 12281-010). .. This custom expression vector replaced the promoter and polyA regions of Gateway pcDNA-DEST47 with the promoter and polyA from the pCAGGS vector ( ) to improve its expression in mammalian neurons.

    Article Title: Concatenation of Human Connexin26 (hCx26) and Human Connexin46 (hCx46) for the Analysis of Heteromeric Gap Junction Hemichannels and Heterotypic Gap Junction Channels
    Article Snippet: .. The purified psDEST47, which is similar to the commercially available pcDNA™-DEST47 vector (#12281010, Thermo Fisher Scientific, Waltham, MA, USA), was used to create a C-terminally GFP-labeled fusion protein via the LR-cloning reaction. .. The vector psGEMHE-GW was used for in vitro transcription to produce the cRNA for the Xenopus oocytes.

    Expressing:

    Article Title: Ca2+ Conduction by an Amino Acid-Gated Ion Channel Related to Glutamate Receptors 1 Conduction by an Amino Acid-Gated Ion Channel Related to Glutamate Receptors 1 [W]
    Article Snippet: .. The resulting PCR fragment was cloned into the pENTR- d entry vector (Invitrogen) followed by a GATEWAY recombination reaction (Invitrogen) into pcDNA-DEST47 mammalian expression vector (pcDEST47-GLR3.4-GFP; for transfection of 293T cells) or the modified pEARLEYGATE 102 plant expression vector ( ) in which the cyan fluorescent protein fragment was replaced with EGFP for enhanced fluorescence to produce the respective 35S:GLR3.4-EGFP . .. The floral dip method was used to transform the glr3.4-1 mutant with the 35S:GLR3.4-EGFP construct.

    Article Title: The Ciliary Protein Arl13b Functions Outside of the Primary Cilium in Shh-Mediated Axon Guidance
    Article Snippet: .. The shRNA/miR/EmGFP sequence was then cloned via attB/attP site recombination (using BP clonase) into the pDONR221 backbone (ThermoFisher, 12536-017), and via attL/attR site recombination (using LR clonase) into a custom expression vector derived from Gateway pcDNA-DEST47 (ThermoFisher Scientific, 12281-010). .. This custom expression vector replaced the promoter and polyA regions of Gateway pcDNA-DEST47 with the promoter and polyA from the pCAGGS vector ( ) to improve its expression in mammalian neurons.

    Sequencing:

    Article Title: The Ciliary Protein Arl13b Functions Outside of the Primary Cilium in Shh-Mediated Axon Guidance
    Article Snippet: .. The shRNA/miR/EmGFP sequence was then cloned via attB/attP site recombination (using BP clonase) into the pDONR221 backbone (ThermoFisher, 12536-017), and via attL/attR site recombination (using LR clonase) into a custom expression vector derived from Gateway pcDNA-DEST47 (ThermoFisher Scientific, 12281-010). .. This custom expression vector replaced the promoter and polyA regions of Gateway pcDNA-DEST47 with the promoter and polyA from the pCAGGS vector ( ) to improve its expression in mammalian neurons.

    Derivative Assay:

    Article Title: The Ciliary Protein Arl13b Functions Outside of the Primary Cilium in Shh-Mediated Axon Guidance
    Article Snippet: .. The shRNA/miR/EmGFP sequence was then cloned via attB/attP site recombination (using BP clonase) into the pDONR221 backbone (ThermoFisher, 12536-017), and via attL/attR site recombination (using LR clonase) into a custom expression vector derived from Gateway pcDNA-DEST47 (ThermoFisher Scientific, 12281-010). .. This custom expression vector replaced the promoter and polyA regions of Gateway pcDNA-DEST47 with the promoter and polyA from the pCAGGS vector ( ) to improve its expression in mammalian neurons.

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  • 94
    Thermo Fisher gateway pcdna dest47
    Gateway Pcdna Dest47, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gateway pcdna dest47/product/Thermo Fisher
    Average 94 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    gateway pcdna dest47 - by Bioz Stars, 2020-09
    94/100 stars
      Buy from Supplier

    92
    Thermo Fisher pcdna dest47 constructs
    Pcdna Dest47 Constructs, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pcdna dest47 constructs/product/Thermo Fisher
    Average 92 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    pcdna dest47 constructs - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

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