gst-affinity chromatography Search Results


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  • 99
    Thermo Fisher gst affinity chromatography
    Gst Affinity Chromatography, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore affinity chromatography gst fusion proteins
    Affinity Chromatography Gst Fusion Proteins, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare gst affinity chromatography
    SDS–PAGE: Purified recombinant <t>VEGFR-2</t> proteins of 11 kDa and 37 kDa. M marker, 1 His-VEGFR-2 protein, 2 <t>GST-VEGFR-2</t> protein
    Gst Affinity Chromatography, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 96/100, based on 420 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare gst affinity column chromatography
    SDS–PAGE: Purified recombinant <t>VEGFR-2</t> proteins of 11 kDa and 37 kDa. M marker, 1 His-VEGFR-2 protein, 2 <t>GST-VEGFR-2</t> protein
    Gst Affinity Column Chromatography, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 99/100, based on 43 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad gst affinity chromatography
    SDS–PAGE: Purified recombinant <t>VEGFR-2</t> proteins of 11 kDa and 37 kDa. M marker, 1 His-VEGFR-2 protein, 2 <t>GST-VEGFR-2</t> protein
    Gst Affinity Chromatography, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 89/100, based on 25 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Stratagene gst affinity chromatography
    Alterations to the RSS sequence can destabilize the <t>RAG</t> SE complex. ( A ) Schematic diagram of a RSS and its adjacent coding sequence. The consensus nucleotide sequence is shown. ( B ) Biochemical end release assay. Purified <t>GST-tagged</t> core RAG proteins cleave a ∼500 bp PCR product at 37°C. Post-cleavage SE complexes are thermally challenged at increasing temperatures and released SEs detected by electrophoresis. ( C ) Representative gels for end release assays. UR, unrearranged substrate; SC, single cleavage; CE, coding ends; SE, signal ends. Numbers above each lane indicate the temperatures reactions were heated to before electrophoresis. Samples treated with proteinase K and SDS are indicated with +pk. ( D ) Quantification of SE release, measured as the combined amount of radioactivity from cleaved products divided by the total amount of radioactivity, from three experiments using two different protein preparations (* P
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    GE Healthcare gst agarose affinity chromatography
    ABA-related gene expression in RhNAC3 -silenced rose petals. A , The putative ABA signaling and downstream rose genes from the ABA-signaling pathway in rose. a, The clone ID from the rose transcriptome database [7] . b, Description of the A. thaliana homolog given by The Arabidopsis Information Resource (TAIR, <t>http://www.arabidopsis.org</t> ). B , qRT-PCR analysis of RhNAC3 -silenced rose petals. The rose cDNAs of TRV and RhNAC3 -silenced (TRV- RhNAC3 ) petals were described in our previous report [26] . Data represent the fold change of each gene by TRV- RhNAC3 relative to the TRV control. RhUbi1 was used as the internal control. Error bars indicate SE ( n = 3). C , Sequences and positions of putative RhNAC3 binding elements used for the EMSA. Probes were derived from the regulatory sequence of three selected ABA-related rose genes. Underlined letters indicate the core sequences of putative NAC protein-binding sites. The sense strands of oligonucleotide probes corresponding to the predicted RhNAC3 binding sites are shown. D , DNA-binding specificity for RhNAC3 with the probes indicated in C . The arrows indicate the positions of protein/DNA complexes and the free probes, respectively. Purified protein (2 µg) was incubated with 0.2 pmol of biotin probe. E , DNA-binding specificity for RhNAC3 with RU03861 . The RU03861 (P3) probe incubated with <t>GST</t> was used as a control, and a 10, 100, and 1000 fold excess of the unlabeled P3 was used for competitive binding.
    Gst Agarose Affinity Chromatography, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 99/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher glutathione s transferase gst affinity chromatography
    ABA-related gene expression in RhNAC3 -silenced rose petals. A , The putative ABA signaling and downstream rose genes from the ABA-signaling pathway in rose. a, The clone ID from the rose transcriptome database [7] . b, Description of the A. thaliana homolog given by The Arabidopsis Information Resource (TAIR, <t>http://www.arabidopsis.org</t> ). B , qRT-PCR analysis of RhNAC3 -silenced rose petals. The rose cDNAs of TRV and RhNAC3 -silenced (TRV- RhNAC3 ) petals were described in our previous report [26] . Data represent the fold change of each gene by TRV- RhNAC3 relative to the TRV control. RhUbi1 was used as the internal control. Error bars indicate SE ( n = 3). C , Sequences and positions of putative RhNAC3 binding elements used for the EMSA. Probes were derived from the regulatory sequence of three selected ABA-related rose genes. Underlined letters indicate the core sequences of putative NAC protein-binding sites. The sense strands of oligonucleotide probes corresponding to the predicted RhNAC3 binding sites are shown. D , DNA-binding specificity for RhNAC3 with the probes indicated in C . The arrows indicate the positions of protein/DNA complexes and the free probes, respectively. Purified protein (2 µg) was incubated with 0.2 pmol of biotin probe. E , DNA-binding specificity for RhNAC3 with RU03861 . The RU03861 (P3) probe incubated with <t>GST</t> was used as a control, and a 10, 100, and 1000 fold excess of the unlabeled P3 was used for competitive binding.
    Glutathione S Transferase Gst Affinity Chromatography, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare glutathione s transferase gst glutathione affinity column chromatography kit
    ABA-related gene expression in RhNAC3 -silenced rose petals. A , The putative ABA signaling and downstream rose genes from the ABA-signaling pathway in rose. a, The clone ID from the rose transcriptome database [7] . b, Description of the A. thaliana homolog given by The Arabidopsis Information Resource (TAIR, <t>http://www.arabidopsis.org</t> ). B , qRT-PCR analysis of RhNAC3 -silenced rose petals. The rose cDNAs of TRV and RhNAC3 -silenced (TRV- RhNAC3 ) petals were described in our previous report [26] . Data represent the fold change of each gene by TRV- RhNAC3 relative to the TRV control. RhUbi1 was used as the internal control. Error bars indicate SE ( n = 3). C , Sequences and positions of putative RhNAC3 binding elements used for the EMSA. Probes were derived from the regulatory sequence of three selected ABA-related rose genes. Underlined letters indicate the core sequences of putative NAC protein-binding sites. The sense strands of oligonucleotide probes corresponding to the predicted RhNAC3 binding sites are shown. D , DNA-binding specificity for RhNAC3 with the probes indicated in C . The arrows indicate the positions of protein/DNA complexes and the free probes, respectively. Purified protein (2 µg) was incubated with 0.2 pmol of biotin probe. E , DNA-binding specificity for RhNAC3 with RU03861 . The RU03861 (P3) probe incubated with <t>GST</t> was used as a control, and a 10, 100, and 1000 fold excess of the unlabeled P3 was used for competitive binding.
    Glutathione S Transferase Gst Glutathione Affinity Column Chromatography Kit, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 95/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare gst affinity chromatographic column
    ABA-related gene expression in RhNAC3 -silenced rose petals. A , The putative ABA signaling and downstream rose genes from the ABA-signaling pathway in rose. a, The clone ID from the rose transcriptome database [7] . b, Description of the A. thaliana homolog given by The Arabidopsis Information Resource (TAIR, <t>http://www.arabidopsis.org</t> ). B , qRT-PCR analysis of RhNAC3 -silenced rose petals. The rose cDNAs of TRV and RhNAC3 -silenced (TRV- RhNAC3 ) petals were described in our previous report [26] . Data represent the fold change of each gene by TRV- RhNAC3 relative to the TRV control. RhUbi1 was used as the internal control. Error bars indicate SE ( n = 3). C , Sequences and positions of putative RhNAC3 binding elements used for the EMSA. Probes were derived from the regulatory sequence of three selected ABA-related rose genes. Underlined letters indicate the core sequences of putative NAC protein-binding sites. The sense strands of oligonucleotide probes corresponding to the predicted RhNAC3 binding sites are shown. D , DNA-binding specificity for RhNAC3 with the probes indicated in C . The arrows indicate the positions of protein/DNA complexes and the free probes, respectively. Purified protein (2 µg) was incubated with 0.2 pmol of biotin probe. E , DNA-binding specificity for RhNAC3 with RU03861 . The RU03861 (P3) probe incubated with <t>GST</t> was used as a control, and a 10, 100, and 1000 fold excess of the unlabeled P3 was used for competitive binding.
    Gst Affinity Chromatographic Column, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 99/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    agios pharma gst affinity column chromatography
    ABA-related gene expression in RhNAC3 -silenced rose petals. A , The putative ABA signaling and downstream rose genes from the ABA-signaling pathway in rose. a, The clone ID from the rose transcriptome database [7] . b, Description of the A. thaliana homolog given by The Arabidopsis Information Resource (TAIR, <t>http://www.arabidopsis.org</t> ). B , qRT-PCR analysis of RhNAC3 -silenced rose petals. The rose cDNAs of TRV and RhNAC3 -silenced (TRV- RhNAC3 ) petals were described in our previous report [26] . Data represent the fold change of each gene by TRV- RhNAC3 relative to the TRV control. RhUbi1 was used as the internal control. Error bars indicate SE ( n = 3). C , Sequences and positions of putative RhNAC3 binding elements used for the EMSA. Probes were derived from the regulatory sequence of three selected ABA-related rose genes. Underlined letters indicate the core sequences of putative NAC protein-binding sites. The sense strands of oligonucleotide probes corresponding to the predicted RhNAC3 binding sites are shown. D , DNA-binding specificity for RhNAC3 with the probes indicated in C . The arrows indicate the positions of protein/DNA complexes and the free probes, respectively. Purified protein (2 µg) was incubated with 0.2 pmol of biotin probe. E , DNA-binding specificity for RhNAC3 with RU03861 . The RU03861 (P3) probe incubated with <t>GST</t> was used as a control, and a 10, 100, and 1000 fold excess of the unlabeled P3 was used for competitive binding.
    Gst Affinity Column Chromatography, supplied by agios pharma, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher gst affinity column chromatography
    ABA-related gene expression in RhNAC3 -silenced rose petals. A , The putative ABA signaling and downstream rose genes from the ABA-signaling pathway in rose. a, The clone ID from the rose transcriptome database [7] . b, Description of the A. thaliana homolog given by The Arabidopsis Information Resource (TAIR, <t>http://www.arabidopsis.org</t> ). B , qRT-PCR analysis of RhNAC3 -silenced rose petals. The rose cDNAs of TRV and RhNAC3 -silenced (TRV- RhNAC3 ) petals were described in our previous report [26] . Data represent the fold change of each gene by TRV- RhNAC3 relative to the TRV control. RhUbi1 was used as the internal control. Error bars indicate SE ( n = 3). C , Sequences and positions of putative RhNAC3 binding elements used for the EMSA. Probes were derived from the regulatory sequence of three selected ABA-related rose genes. Underlined letters indicate the core sequences of putative NAC protein-binding sites. The sense strands of oligonucleotide probes corresponding to the predicted RhNAC3 binding sites are shown. D , DNA-binding specificity for RhNAC3 with the probes indicated in C . The arrows indicate the positions of protein/DNA complexes and the free probes, respectively. Purified protein (2 µg) was incubated with 0.2 pmol of biotin probe. E , DNA-binding specificity for RhNAC3 with RU03861 . The RU03861 (P3) probe incubated with <t>GST</t> was used as a control, and a 10, 100, and 1000 fold excess of the unlabeled P3 was used for competitive binding.
    Gst Affinity Column Chromatography, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare gst bead affinity chromatography
    ABA-related gene expression in RhNAC3 -silenced rose petals. A , The putative ABA signaling and downstream rose genes from the ABA-signaling pathway in rose. a, The clone ID from the rose transcriptome database [7] . b, Description of the A. thaliana homolog given by The Arabidopsis Information Resource (TAIR, <t>http://www.arabidopsis.org</t> ). B , qRT-PCR analysis of RhNAC3 -silenced rose petals. The rose cDNAs of TRV and RhNAC3 -silenced (TRV- RhNAC3 ) petals were described in our previous report [26] . Data represent the fold change of each gene by TRV- RhNAC3 relative to the TRV control. RhUbi1 was used as the internal control. Error bars indicate SE ( n = 3). C , Sequences and positions of putative RhNAC3 binding elements used for the EMSA. Probes were derived from the regulatory sequence of three selected ABA-related rose genes. Underlined letters indicate the core sequences of putative NAC protein-binding sites. The sense strands of oligonucleotide probes corresponding to the predicted RhNAC3 binding sites are shown. D , DNA-binding specificity for RhNAC3 with the probes indicated in C . The arrows indicate the positions of protein/DNA complexes and the free probes, respectively. Purified protein (2 µg) was incubated with 0.2 pmol of biotin probe. E , DNA-binding specificity for RhNAC3 with RU03861 . The RU03861 (P3) probe incubated with <t>GST</t> was used as a control, and a 10, 100, and 1000 fold excess of the unlabeled P3 was used for competitive binding.
    Gst Bead Affinity Chromatography, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 99/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare gst affinity chromatography column
    ABA-related gene expression in RhNAC3 -silenced rose petals. A , The putative ABA signaling and downstream rose genes from the ABA-signaling pathway in rose. a, The clone ID from the rose transcriptome database [7] . b, Description of the A. thaliana homolog given by The Arabidopsis Information Resource (TAIR, <t>http://www.arabidopsis.org</t> ). B , qRT-PCR analysis of RhNAC3 -silenced rose petals. The rose cDNAs of TRV and RhNAC3 -silenced (TRV- RhNAC3 ) petals were described in our previous report [26] . Data represent the fold change of each gene by TRV- RhNAC3 relative to the TRV control. RhUbi1 was used as the internal control. Error bars indicate SE ( n = 3). C , Sequences and positions of putative RhNAC3 binding elements used for the EMSA. Probes were derived from the regulatory sequence of three selected ABA-related rose genes. Underlined letters indicate the core sequences of putative NAC protein-binding sites. The sense strands of oligonucleotide probes corresponding to the predicted RhNAC3 binding sites are shown. D , DNA-binding specificity for RhNAC3 with the probes indicated in C . The arrows indicate the positions of protein/DNA complexes and the free probes, respectively. Purified protein (2 µg) was incubated with 0.2 pmol of biotin probe. E , DNA-binding specificity for RhNAC3 with RU03861 . The RU03861 (P3) probe incubated with <t>GST</t> was used as a control, and a 10, 100, and 1000 fold excess of the unlabeled P3 was used for competitive binding.
    Gst Affinity Chromatography Column, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 86/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare gst affinity chromatography columns
    ABA-related gene expression in RhNAC3 -silenced rose petals. A , The putative ABA signaling and downstream rose genes from the ABA-signaling pathway in rose. a, The clone ID from the rose transcriptome database [7] . b, Description of the A. thaliana homolog given by The Arabidopsis Information Resource (TAIR, <t>http://www.arabidopsis.org</t> ). B , qRT-PCR analysis of RhNAC3 -silenced rose petals. The rose cDNAs of TRV and RhNAC3 -silenced (TRV- RhNAC3 ) petals were described in our previous report [26] . Data represent the fold change of each gene by TRV- RhNAC3 relative to the TRV control. RhUbi1 was used as the internal control. Error bars indicate SE ( n = 3). C , Sequences and positions of putative RhNAC3 binding elements used for the EMSA. Probes were derived from the regulatory sequence of three selected ABA-related rose genes. Underlined letters indicate the core sequences of putative NAC protein-binding sites. The sense strands of oligonucleotide probes corresponding to the predicted RhNAC3 binding sites are shown. D , DNA-binding specificity for RhNAC3 with the probes indicated in C . The arrows indicate the positions of protein/DNA complexes and the free probes, respectively. Purified protein (2 µg) was incubated with 0.2 pmol of biotin probe. E , DNA-binding specificity for RhNAC3 with RU03861 . The RU03861 (P3) probe incubated with <t>GST</t> was used as a control, and a 10, 100, and 1000 fold excess of the unlabeled P3 was used for competitive binding.
    Gst Affinity Chromatography Columns, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 99/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher gst affinity chromatography columns
    ABA-related gene expression in RhNAC3 -silenced rose petals. A , The putative ABA signaling and downstream rose genes from the ABA-signaling pathway in rose. a, The clone ID from the rose transcriptome database [7] . b, Description of the A. thaliana homolog given by The Arabidopsis Information Resource (TAIR, <t>http://www.arabidopsis.org</t> ). B , qRT-PCR analysis of RhNAC3 -silenced rose petals. The rose cDNAs of TRV and RhNAC3 -silenced (TRV- RhNAC3 ) petals were described in our previous report [26] . Data represent the fold change of each gene by TRV- RhNAC3 relative to the TRV control. RhUbi1 was used as the internal control. Error bars indicate SE ( n = 3). C , Sequences and positions of putative RhNAC3 binding elements used for the EMSA. Probes were derived from the regulatory sequence of three selected ABA-related rose genes. Underlined letters indicate the core sequences of putative NAC protein-binding sites. The sense strands of oligonucleotide probes corresponding to the predicted RhNAC3 binding sites are shown. D , DNA-binding specificity for RhNAC3 with the probes indicated in C . The arrows indicate the positions of protein/DNA complexes and the free probes, respectively. Purified protein (2 µg) was incubated with 0.2 pmol of biotin probe. E , DNA-binding specificity for RhNAC3 with RU03861 . The RU03861 (P3) probe incubated with <t>GST</t> was used as a control, and a 10, 100, and 1000 fold excess of the unlabeled P3 was used for competitive binding.
    Gst Affinity Chromatography Columns, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ABA-related gene expression in RhNAC3 -silenced rose petals. A , The putative ABA signaling and downstream rose genes from the ABA-signaling pathway in rose. a, The clone ID from the rose transcriptome database [7] . b, Description of the A. thaliana homolog given by The Arabidopsis Information Resource (TAIR, <t>http://www.arabidopsis.org</t> ). B , qRT-PCR analysis of RhNAC3 -silenced rose petals. The rose cDNAs of TRV and RhNAC3 -silenced (TRV- RhNAC3 ) petals were described in our previous report [26] . Data represent the fold change of each gene by TRV- RhNAC3 relative to the TRV control. RhUbi1 was used as the internal control. Error bars indicate SE ( n = 3). C , Sequences and positions of putative RhNAC3 binding elements used for the EMSA. Probes were derived from the regulatory sequence of three selected ABA-related rose genes. Underlined letters indicate the core sequences of putative NAC protein-binding sites. The sense strands of oligonucleotide probes corresponding to the predicted RhNAC3 binding sites are shown. D , DNA-binding specificity for RhNAC3 with the probes indicated in C . The arrows indicate the positions of protein/DNA complexes and the free probes, respectively. Purified protein (2 µg) was incubated with 0.2 pmol of biotin probe. E , DNA-binding specificity for RhNAC3 with RU03861 . The RU03861 (P3) probe incubated with <t>GST</t> was used as a control, and a 10, 100, and 1000 fold excess of the unlabeled P3 was used for competitive binding.
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    Millipore gst sepharose sigma aldrich affinity chromatography
    ABA-related gene expression in RhNAC3 -silenced rose petals. A , The putative ABA signaling and downstream rose genes from the ABA-signaling pathway in rose. a, The clone ID from the rose transcriptome database [7] . b, Description of the A. thaliana homolog given by The Arabidopsis Information Resource (TAIR, <t>http://www.arabidopsis.org</t> ). B , qRT-PCR analysis of RhNAC3 -silenced rose petals. The rose cDNAs of TRV and RhNAC3 -silenced (TRV- RhNAC3 ) petals were described in our previous report [26] . Data represent the fold change of each gene by TRV- RhNAC3 relative to the TRV control. RhUbi1 was used as the internal control. Error bars indicate SE ( n = 3). C , Sequences and positions of putative RhNAC3 binding elements used for the EMSA. Probes were derived from the regulatory sequence of three selected ABA-related rose genes. Underlined letters indicate the core sequences of putative NAC protein-binding sites. The sense strands of oligonucleotide probes corresponding to the predicted RhNAC3 binding sites are shown. D , DNA-binding specificity for RhNAC3 with the probes indicated in C . The arrows indicate the positions of protein/DNA complexes and the free probes, respectively. Purified protein (2 µg) was incubated with 0.2 pmol of biotin probe. E , DNA-binding specificity for RhNAC3 with RU03861 . The RU03861 (P3) probe incubated with <t>GST</t> was used as a control, and a 10, 100, and 1000 fold excess of the unlabeled P3 was used for competitive binding.
    Gst Sepharose Sigma Aldrich Affinity Chromatography, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    SDS–PAGE: Purified recombinant VEGFR-2 proteins of 11 kDa and 37 kDa. M marker, 1 His-VEGFR-2 protein, 2 GST-VEGFR-2 protein

    Journal: Cytotechnology

    Article Title: A high-affinity human/mouse cross-reactive monoclonal antibody, specific for VEGFR-2 linear and conformational epitopes

    doi: 10.1007/s10616-010-9262-4

    Figure Lengend Snippet: SDS–PAGE: Purified recombinant VEGFR-2 proteins of 11 kDa and 37 kDa. M marker, 1 His-VEGFR-2 protein, 2 GST-VEGFR-2 protein

    Article Snippet: His-tagged VEGFR-2 (His-VEGFR-2) protein and glutathione S-transferase-fusion VEGFR-2 (GST-VEGFR-2) protein in the bacilli ultrasonic supernatant were purified by His-tag and GST-trap affinity chromatography (GE, USA) using the GE purification system.

    Techniques: SDS Page, Purification, Recombinant, Marker

    Alterations to the RSS sequence can destabilize the RAG SE complex. ( A ) Schematic diagram of a RSS and its adjacent coding sequence. The consensus nucleotide sequence is shown. ( B ) Biochemical end release assay. Purified GST-tagged core RAG proteins cleave a ∼500 bp PCR product at 37°C. Post-cleavage SE complexes are thermally challenged at increasing temperatures and released SEs detected by electrophoresis. ( C ) Representative gels for end release assays. UR, unrearranged substrate; SC, single cleavage; CE, coding ends; SE, signal ends. Numbers above each lane indicate the temperatures reactions were heated to before electrophoresis. Samples treated with proteinase K and SDS are indicated with +pk. ( D ) Quantification of SE release, measured as the combined amount of radioactivity from cleaved products divided by the total amount of radioactivity, from three experiments using two different protein preparations (* P

    Journal: Nucleic Acids Research

    Article Title: Non-consensus heptamer sequences destabilize the RAG post-cleavage complex, making ends available to alternative DNA repair pathways

    doi: 10.1093/nar/gkp1252

    Figure Lengend Snippet: Alterations to the RSS sequence can destabilize the RAG SE complex. ( A ) Schematic diagram of a RSS and its adjacent coding sequence. The consensus nucleotide sequence is shown. ( B ) Biochemical end release assay. Purified GST-tagged core RAG proteins cleave a ∼500 bp PCR product at 37°C. Post-cleavage SE complexes are thermally challenged at increasing temperatures and released SEs detected by electrophoresis. ( C ) Representative gels for end release assays. UR, unrearranged substrate; SC, single cleavage; CE, coding ends; SE, signal ends. Numbers above each lane indicate the temperatures reactions were heated to before electrophoresis. Samples treated with proteinase K and SDS are indicated with +pk. ( D ) Quantification of SE release, measured as the combined amount of radioactivity from cleaved products divided by the total amount of radioactivity, from three experiments using two different protein preparations (* P

    Article Snippet: GST-tagged truncated RAG proteins were purified from RMP41 fibroblasts by GST affinity chromatography ( ) using GST affinity resin (Stratagene).

    Techniques: Sequencing, Release Assay, Purification, Polymerase Chain Reaction, Electrophoresis, Radioactivity

    ABA-related gene expression in RhNAC3 -silenced rose petals. A , The putative ABA signaling and downstream rose genes from the ABA-signaling pathway in rose. a, The clone ID from the rose transcriptome database [7] . b, Description of the A. thaliana homolog given by The Arabidopsis Information Resource (TAIR, http://www.arabidopsis.org ). B , qRT-PCR analysis of RhNAC3 -silenced rose petals. The rose cDNAs of TRV and RhNAC3 -silenced (TRV- RhNAC3 ) petals were described in our previous report [26] . Data represent the fold change of each gene by TRV- RhNAC3 relative to the TRV control. RhUbi1 was used as the internal control. Error bars indicate SE ( n = 3). C , Sequences and positions of putative RhNAC3 binding elements used for the EMSA. Probes were derived from the regulatory sequence of three selected ABA-related rose genes. Underlined letters indicate the core sequences of putative NAC protein-binding sites. The sense strands of oligonucleotide probes corresponding to the predicted RhNAC3 binding sites are shown. D , DNA-binding specificity for RhNAC3 with the probes indicated in C . The arrows indicate the positions of protein/DNA complexes and the free probes, respectively. Purified protein (2 µg) was incubated with 0.2 pmol of biotin probe. E , DNA-binding specificity for RhNAC3 with RU03861 . The RU03861 (P3) probe incubated with GST was used as a control, and a 10, 100, and 1000 fold excess of the unlabeled P3 was used for competitive binding.

    Journal: PLoS ONE

    Article Title: The Rose (Rosa hybrida) NAC Transcription Factor 3 Gene, RhNAC3, Involved in ABA Signaling Pathway Both in Rose and Arabidopsis

    doi: 10.1371/journal.pone.0109415

    Figure Lengend Snippet: ABA-related gene expression in RhNAC3 -silenced rose petals. A , The putative ABA signaling and downstream rose genes from the ABA-signaling pathway in rose. a, The clone ID from the rose transcriptome database [7] . b, Description of the A. thaliana homolog given by The Arabidopsis Information Resource (TAIR, http://www.arabidopsis.org ). B , qRT-PCR analysis of RhNAC3 -silenced rose petals. The rose cDNAs of TRV and RhNAC3 -silenced (TRV- RhNAC3 ) petals were described in our previous report [26] . Data represent the fold change of each gene by TRV- RhNAC3 relative to the TRV control. RhUbi1 was used as the internal control. Error bars indicate SE ( n = 3). C , Sequences and positions of putative RhNAC3 binding elements used for the EMSA. Probes were derived from the regulatory sequence of three selected ABA-related rose genes. Underlined letters indicate the core sequences of putative NAC protein-binding sites. The sense strands of oligonucleotide probes corresponding to the predicted RhNAC3 binding sites are shown. D , DNA-binding specificity for RhNAC3 with the probes indicated in C . The arrows indicate the positions of protein/DNA complexes and the free probes, respectively. Purified protein (2 µg) was incubated with 0.2 pmol of biotin probe. E , DNA-binding specificity for RhNAC3 with RU03861 . The RU03861 (P3) probe incubated with GST was used as a control, and a 10, 100, and 1000 fold excess of the unlabeled P3 was used for competitive binding.

    Article Snippet: The fusion protein was induced by 0.2 mM isopropyl β-D-1-thiogalactopyranoside (IPTG), and the cells E. coli incubated at 28°C for a further 6 h. The recombinant protein was purified by GST-agarose affinity chromatography (GE Healthcare, http://www.gehealthcare.com/ ).

    Techniques: Expressing, Quantitative RT-PCR, Binding Assay, Derivative Assay, Sequencing, Protein Binding, Purification, Incubation