topoisomerase activated pcr 2 1 topo vector  (Thermo Fisher)


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    Thermo Fisher topoisomerase activated pcr 2 1 topo vector
    Topoisomerase Activated Pcr 2 1 Topo Vector, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/topoisomerase activated pcr 2 1 topo vector/product/Thermo Fisher
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    topoisomerase activated pcr 2 1 topo vector - by Bioz Stars, 2020-08
    85/100 stars

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    Related Articles

    Polymerase Chain Reaction:

    Article Title: Purification, crystallization and preliminary X-ray analysis of a thermostable direct haemolysin from Grimontia hollisae
    Article Snippet: .. The recombinant pTOPO- Gh-tdh expression vector was obtained by direct insertion of the Taq polymerase-amplified PCR product of the full-length Gh-tdh gene from G. hollisae (ATCC 33564) into topoisomerase-activated pCR 2.1-TOPO vector (Invitrogen, Carlsbad, California, USA) according to the manufacturer’s protocol. .. The recombinant plasmid was sequenced using an ABI PRISM 3100 genetic analyzer to ensure its fidelity in the subsequent protein-expression and purification experiments.

    Recombinant:

    Article Title: Purification, crystallization and preliminary X-ray analysis of a thermostable direct haemolysin from Grimontia hollisae
    Article Snippet: .. The recombinant pTOPO- Gh-tdh expression vector was obtained by direct insertion of the Taq polymerase-amplified PCR product of the full-length Gh-tdh gene from G. hollisae (ATCC 33564) into topoisomerase-activated pCR 2.1-TOPO vector (Invitrogen, Carlsbad, California, USA) according to the manufacturer’s protocol. .. The recombinant plasmid was sequenced using an ABI PRISM 3100 genetic analyzer to ensure its fidelity in the subsequent protein-expression and purification experiments.

    Expressing:

    Article Title: Purification, crystallization and preliminary X-ray analysis of a thermostable direct haemolysin from Grimontia hollisae
    Article Snippet: .. The recombinant pTOPO- Gh-tdh expression vector was obtained by direct insertion of the Taq polymerase-amplified PCR product of the full-length Gh-tdh gene from G. hollisae (ATCC 33564) into topoisomerase-activated pCR 2.1-TOPO vector (Invitrogen, Carlsbad, California, USA) according to the manufacturer’s protocol. .. The recombinant plasmid was sequenced using an ABI PRISM 3100 genetic analyzer to ensure its fidelity in the subsequent protein-expression and purification experiments.

    Plasmid Preparation:

    Article Title: Purification, crystallization and preliminary X-ray analysis of a thermostable direct haemolysin from Grimontia hollisae
    Article Snippet: .. The recombinant pTOPO- Gh-tdh expression vector was obtained by direct insertion of the Taq polymerase-amplified PCR product of the full-length Gh-tdh gene from G. hollisae (ATCC 33564) into topoisomerase-activated pCR 2.1-TOPO vector (Invitrogen, Carlsbad, California, USA) according to the manufacturer’s protocol. .. The recombinant plasmid was sequenced using an ABI PRISM 3100 genetic analyzer to ensure its fidelity in the subsequent protein-expression and purification experiments.

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    Thermo Fisher gene exp topors mm00506480 m1
    Deletion analysis of the S1CD-interacting domain of <t>Topors.</t> ( A ) Model of murine Topors protein domains, with the sequence corresponding to the S1CD-interacting prey 36 sequence identified by a black box. ( B ) Deletional analysis of the S1CD-binding sequence of Topors. Yeast with either LexA-S1CD or LexA-Lamin bait plasmid were transformed with either prey plasmid for the expression of complete S1CD binding sequence (Topors 398–516), or with prey plasmid expressing deletion mutants of Topors 398–516. After selection of co-transformants, interacting bait-prey pairs were identified by assay of β-galactosidase activity. Yeast two-hybrid analysis of Topors deletion mutants identifies residues 493–510 as critical for the interaction.
    Gene Exp Topors Mm00506480 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gene exp topors mm00506480 m1/product/Thermo Fisher
    Average 85 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    gene exp topors mm00506480 m1 - by Bioz Stars, 2020-08
    85/100 stars
      Buy from Supplier

    99
    Thermo Fisher gene exp top1 hs00243257 m1
    Deletion analysis of the S1CD-interacting domain of <t>Topors.</t> ( A ) Model of murine Topors protein domains, with the sequence corresponding to the S1CD-interacting prey 36 sequence identified by a black box. ( B ) Deletional analysis of the S1CD-binding sequence of Topors. Yeast with either LexA-S1CD or LexA-Lamin bait plasmid were transformed with either prey plasmid for the expression of complete S1CD binding sequence (Topors 398–516), or with prey plasmid expressing deletion mutants of Topors 398–516. After selection of co-transformants, interacting bait-prey pairs were identified by assay of β-galactosidase activity. Yeast two-hybrid analysis of Topors deletion mutants identifies residues 493–510 as critical for the interaction.
    Gene Exp Top1 Hs00243257 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gene exp top1 hs00243257 m1/product/Thermo Fisher
    Average 99 stars, based on 9 article reviews
    Price from $9.99 to $1999.99
    gene exp top1 hs00243257 m1 - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

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    Deletion analysis of the S1CD-interacting domain of Topors. ( A ) Model of murine Topors protein domains, with the sequence corresponding to the S1CD-interacting prey 36 sequence identified by a black box. ( B ) Deletional analysis of the S1CD-binding sequence of Topors. Yeast with either LexA-S1CD or LexA-Lamin bait plasmid were transformed with either prey plasmid for the expression of complete S1CD binding sequence (Topors 398–516), or with prey plasmid expressing deletion mutants of Topors 398–516. After selection of co-transformants, interacting bait-prey pairs were identified by assay of β-galactosidase activity. Yeast two-hybrid analysis of Topors deletion mutants identifies residues 493–510 as critical for the interaction.

    Journal: PLoS ONE

    Article Title: Inhibition of PDGF-B Induction and Cell Growth by Syndecan-1 Involves the Ubiquitin and SUMO-1 Ligase, Topors

    doi: 10.1371/journal.pone.0043701

    Figure Lengend Snippet: Deletion analysis of the S1CD-interacting domain of Topors. ( A ) Model of murine Topors protein domains, with the sequence corresponding to the S1CD-interacting prey 36 sequence identified by a black box. ( B ) Deletional analysis of the S1CD-binding sequence of Topors. Yeast with either LexA-S1CD or LexA-Lamin bait plasmid were transformed with either prey plasmid for the expression of complete S1CD binding sequence (Topors 398–516), or with prey plasmid expressing deletion mutants of Topors 398–516. After selection of co-transformants, interacting bait-prey pairs were identified by assay of β-galactosidase activity. Yeast two-hybrid analysis of Topors deletion mutants identifies residues 493–510 as critical for the interaction.

    Article Snippet: Separate samples were used to extract RNA for qPCR for Topors that was done using the Applied Biosystems Taqman probe for Topors (Assay# Mm00506480_m1).

    Techniques: Sequencing, Binding Assay, Plasmid Preparation, Transformation Assay, Expressing, Selection, Activity Assay

    Co-immune precipitation of Sdc-1 and Topors from lysates of NMuMG cells. ( A ) Total cell lysates prepared from NMuMG cells were immune precipitated (IP) with non-immune rat IgG (lane 1) or rat anti-Sdc-1 antibody (lane 2 and 3) and then western blotted for Sdc-1 (lanes 1–2), and after stripping, re-probed with anti-Topors (lanes 3). Total cell lysate (lane 4) was included as a control for anti-Topors immunoreactivity. Arrows mark major co-precipitated Topors bands (110 and 175 kDa). ( B ) Non-immune chicken IgY-preadsorbed total cell lysates of NMuMG cells were immunoprecipitated with chicken anti-Topors, and immune precipitates were heparinase- and chondroitin ABC lyase-digested prior to SDS-PAGE and western blotting. Lysate (lane 1) and Topors immunoprecipitates (lane 2 and 3) that were probed on western blots for Topors (lanes 1 and 2) had Topors bands (110 and 175 kDa bands, double arrow), or for Sdc-1 (lane 3), which gave a band of ∼85 kDa (large arrow, lane 3 and 4), similar in size to a band present in a Sdc-1-positive control (lane 4), prepared by ion-exchange chromatography from the medium of NMuMG cultures. Lower MW heavy bands represent chicken IgY (NChIgY) from the immune precipitate (arrow). Lanes 1–3 are from the same blot with lane 3 representing a stripped and re-probed lane of the Topors immune precipitate run in parallel to that shown in lane 2. Lane 4 represents an adjacent lane from the same blot, at a longer exposure time.

    Journal: PLoS ONE

    Article Title: Inhibition of PDGF-B Induction and Cell Growth by Syndecan-1 Involves the Ubiquitin and SUMO-1 Ligase, Topors

    doi: 10.1371/journal.pone.0043701

    Figure Lengend Snippet: Co-immune precipitation of Sdc-1 and Topors from lysates of NMuMG cells. ( A ) Total cell lysates prepared from NMuMG cells were immune precipitated (IP) with non-immune rat IgG (lane 1) or rat anti-Sdc-1 antibody (lane 2 and 3) and then western blotted for Sdc-1 (lanes 1–2), and after stripping, re-probed with anti-Topors (lanes 3). Total cell lysate (lane 4) was included as a control for anti-Topors immunoreactivity. Arrows mark major co-precipitated Topors bands (110 and 175 kDa). ( B ) Non-immune chicken IgY-preadsorbed total cell lysates of NMuMG cells were immunoprecipitated with chicken anti-Topors, and immune precipitates were heparinase- and chondroitin ABC lyase-digested prior to SDS-PAGE and western blotting. Lysate (lane 1) and Topors immunoprecipitates (lane 2 and 3) that were probed on western blots for Topors (lanes 1 and 2) had Topors bands (110 and 175 kDa bands, double arrow), or for Sdc-1 (lane 3), which gave a band of ∼85 kDa (large arrow, lane 3 and 4), similar in size to a band present in a Sdc-1-positive control (lane 4), prepared by ion-exchange chromatography from the medium of NMuMG cultures. Lower MW heavy bands represent chicken IgY (NChIgY) from the immune precipitate (arrow). Lanes 1–3 are from the same blot with lane 3 representing a stripped and re-probed lane of the Topors immune precipitate run in parallel to that shown in lane 2. Lane 4 represents an adjacent lane from the same blot, at a longer exposure time.

    Article Snippet: Separate samples were used to extract RNA for qPCR for Topors that was done using the Applied Biosystems Taqman probe for Topors (Assay# Mm00506480_m1).

    Techniques: Western Blot, Stripping Membranes, Immunoprecipitation, SDS Page, Positive Control, Ion Exchange Chromatography

    Affinity capture and western blotting of Topors. ( A ) Western blot of NMuMG cell detergent lysate (supernate) and detergent-insoluble material (pellet) probed with chicken anti-Topors antibody. Two major bands (∼110 and ∼175 kDa (arrowheads)) are apparent. The larger band is prevalent in detergent-insoluble fractions, which contain relatively more histone proteins (lower blot), consistent with enhanced presence of nuclear proteins. ( B ) Capture of endogenous Topors from cell lysates by GST-S1CD. GST-S1CD or GST alone were adsorbed to glutathione-agarose beads and incubated with detergent lysates of 3T3 cells. Anti-Topors-reactive bands captured by GST-S1CD were detected in lysates of 3T3 cells that overexpress Topors (first lane, arrows), and were similar in size to the major bands seen in western blots of whole cell lysates. Similar bands were not captured by GST alone (lane 2), nor are they present in the GST-S1CD reagent (lane 3). Non-specific bands are seen at ∼55 kDa. ( C ) Co-precipitation of Topors with polyhistidine-tagged Sdc-1 isolated by metal-ion affinity chromatography. Metal ion-affinity gel bound material was isolated from detergent extracts of cells that express polyhistidine-tagged Sdc-1. Parallel samples were run on SDS-PAGE, either directly or after heparinase and chondroitinase digestion and transferred to nitrocellulose. Parallel blots were probed with chicken anti-Topors or, for digested samples, with anti Sdc-1.

    Journal: PLoS ONE

    Article Title: Inhibition of PDGF-B Induction and Cell Growth by Syndecan-1 Involves the Ubiquitin and SUMO-1 Ligase, Topors

    doi: 10.1371/journal.pone.0043701

    Figure Lengend Snippet: Affinity capture and western blotting of Topors. ( A ) Western blot of NMuMG cell detergent lysate (supernate) and detergent-insoluble material (pellet) probed with chicken anti-Topors antibody. Two major bands (∼110 and ∼175 kDa (arrowheads)) are apparent. The larger band is prevalent in detergent-insoluble fractions, which contain relatively more histone proteins (lower blot), consistent with enhanced presence of nuclear proteins. ( B ) Capture of endogenous Topors from cell lysates by GST-S1CD. GST-S1CD or GST alone were adsorbed to glutathione-agarose beads and incubated with detergent lysates of 3T3 cells. Anti-Topors-reactive bands captured by GST-S1CD were detected in lysates of 3T3 cells that overexpress Topors (first lane, arrows), and were similar in size to the major bands seen in western blots of whole cell lysates. Similar bands were not captured by GST alone (lane 2), nor are they present in the GST-S1CD reagent (lane 3). Non-specific bands are seen at ∼55 kDa. ( C ) Co-precipitation of Topors with polyhistidine-tagged Sdc-1 isolated by metal-ion affinity chromatography. Metal ion-affinity gel bound material was isolated from detergent extracts of cells that express polyhistidine-tagged Sdc-1. Parallel samples were run on SDS-PAGE, either directly or after heparinase and chondroitinase digestion and transferred to nitrocellulose. Parallel blots were probed with chicken anti-Topors or, for digested samples, with anti Sdc-1.

    Article Snippet: Separate samples were used to extract RNA for qPCR for Topors that was done using the Applied Biosystems Taqman probe for Topors (Assay# Mm00506480_m1).

    Techniques: Western Blot, Incubation, Isolation, Affinity Chromatography, SDS Page

    The effect of siRNA-mediated Topors knockdown on cell proliferation and PDGF-B chain expression by wild-type and Sdc-1 null SMCs. ( A ) The effect of siRNA to Topors (closed bars) or a scrambled control (open bars) on [ 3 H]-thymidine incorporation mediated by thrombin in wild-type and Sdc-1 null SMCs. Values are the mean ± SEM of four experiments expressed as the fold of wild-type scrambled siRNA control. ( B ) The same data as in section (A) except that data is expressed as the fold of its appropriate control (either the scrambled control or the siRNA to Topors of either wild-type or Sdc-1 null). ( C ) The effect of siRNA to Topors (closed bars) or a scrambled control (open bars) on PDGF-B chain mRNA mediated by thrombin in wild-type and Sdc-1 null SMCs. Values are the mean ± SEM of four experiments for thrombin expressed as the fold of wild-type scrambled siRNA control. * P

    Journal: PLoS ONE

    Article Title: Inhibition of PDGF-B Induction and Cell Growth by Syndecan-1 Involves the Ubiquitin and SUMO-1 Ligase, Topors

    doi: 10.1371/journal.pone.0043701

    Figure Lengend Snippet: The effect of siRNA-mediated Topors knockdown on cell proliferation and PDGF-B chain expression by wild-type and Sdc-1 null SMCs. ( A ) The effect of siRNA to Topors (closed bars) or a scrambled control (open bars) on [ 3 H]-thymidine incorporation mediated by thrombin in wild-type and Sdc-1 null SMCs. Values are the mean ± SEM of four experiments expressed as the fold of wild-type scrambled siRNA control. ( B ) The same data as in section (A) except that data is expressed as the fold of its appropriate control (either the scrambled control or the siRNA to Topors of either wild-type or Sdc-1 null). ( C ) The effect of siRNA to Topors (closed bars) or a scrambled control (open bars) on PDGF-B chain mRNA mediated by thrombin in wild-type and Sdc-1 null SMCs. Values are the mean ± SEM of four experiments for thrombin expressed as the fold of wild-type scrambled siRNA control. * P

    Article Snippet: Separate samples were used to extract RNA for qPCR for Topors that was done using the Applied Biosystems Taqman probe for Topors (Assay# Mm00506480_m1).

    Techniques: Expressing

    Immunolocalization of Topors and Sdc-1. ( A ) Immunofluorescent staining of confluent NMuMG cells demonstrates that Topors is present in a punctate pattern in nuclei (e.g., arrowhead), consistent with previous reports of nuclear localization. In addition, Topors immunoreactivity is present at cell boundaries (arrows) as well as heavily present in the cytoplasm of cells undergoing mitosis (open arrow). Bar = 20 μm. ( B ) Immunofluorescent images of a single field of confluent NMuMG cells immunostained for mouse Sdc-1 (green, left) and Topors (red, right), with a merged image presented in the middle panel. Bar = 20 μm. ( C ) Immunofluorescent staining of dorsal epidermis of an e13.5 mouse embryo for Sdc-1 (red, left) and Topors (green, right). The merged imaged (center) includes a nuclear stain (DAPI, blue). Bar = 50 μm.

    Journal: PLoS ONE

    Article Title: Inhibition of PDGF-B Induction and Cell Growth by Syndecan-1 Involves the Ubiquitin and SUMO-1 Ligase, Topors

    doi: 10.1371/journal.pone.0043701

    Figure Lengend Snippet: Immunolocalization of Topors and Sdc-1. ( A ) Immunofluorescent staining of confluent NMuMG cells demonstrates that Topors is present in a punctate pattern in nuclei (e.g., arrowhead), consistent with previous reports of nuclear localization. In addition, Topors immunoreactivity is present at cell boundaries (arrows) as well as heavily present in the cytoplasm of cells undergoing mitosis (open arrow). Bar = 20 μm. ( B ) Immunofluorescent images of a single field of confluent NMuMG cells immunostained for mouse Sdc-1 (green, left) and Topors (red, right), with a merged image presented in the middle panel. Bar = 20 μm. ( C ) Immunofluorescent staining of dorsal epidermis of an e13.5 mouse embryo for Sdc-1 (red, left) and Topors (green, right). The merged imaged (center) includes a nuclear stain (DAPI, blue). Bar = 50 μm.

    Article Snippet: Separate samples were used to extract RNA for qPCR for Topors that was done using the Applied Biosystems Taqman probe for Topors (Assay# Mm00506480_m1).

    Techniques: Staining