shuffle strain c3030  (New England Biolabs)


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    New England Biolabs shuffle strain c3030
    Cytoplamic expression of proTHI-TRX fusion proteins in strain <t>C3030.</t> For all fusion proteins, 1 ml of cell culture was pelleted and dissolved in 100 μl sample buffer. 10 μl from this extract and an equivalent amount for total soluble cytoplasmic fractions were separated on Tricine/SDS gels. (M) Protein marker, ( 1 ) un-induced crude extract, ( 2 ) induced crude extract, ( 3 ) total soluble fraction taken after cell lysis by sonication, ( 4 ) insoluble fraction after sonication. Red stars indicate the 25 kDa fusion protein
    Shuffle Strain C3030, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 78/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/shuffle strain c3030/product/New England Biolabs
    Average 78 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    shuffle strain c3030 - by Bioz Stars, 2019-07
    78/100 stars

    Images

    1) Product Images from "Comparison of periplasmic and intracellular expression of Arabidopsis thionin proproteins in E. coli"

    Article Title: Comparison of periplasmic and intracellular expression of Arabidopsis thionin proproteins in E. coli

    Journal: Biotechnology Letters

    doi: 10.1007/s10529-013-1180-z

    Cytoplamic expression of proTHI-TRX fusion proteins in strain C3030. For all fusion proteins, 1 ml of cell culture was pelleted and dissolved in 100 μl sample buffer. 10 μl from this extract and an equivalent amount for total soluble cytoplasmic fractions were separated on Tricine/SDS gels. (M) Protein marker, ( 1 ) un-induced crude extract, ( 2 ) induced crude extract, ( 3 ) total soluble fraction taken after cell lysis by sonication, ( 4 ) insoluble fraction after sonication. Red stars indicate the 25 kDa fusion protein
    Figure Legend Snippet: Cytoplamic expression of proTHI-TRX fusion proteins in strain C3030. For all fusion proteins, 1 ml of cell culture was pelleted and dissolved in 100 μl sample buffer. 10 μl from this extract and an equivalent amount for total soluble cytoplasmic fractions were separated on Tricine/SDS gels. (M) Protein marker, ( 1 ) un-induced crude extract, ( 2 ) induced crude extract, ( 3 ) total soluble fraction taken after cell lysis by sonication, ( 4 ) insoluble fraction after sonication. Red stars indicate the 25 kDa fusion protein

    Techniques Used: Expressing, Cell Culture, Marker, Lysis, Sonication

    Comparison of Ni–NTA purified proTHI-TRX fusion proteins from strain C3030. a Coomassie Brilliant Blue staining. b Western blot with anti-His tag antibody. Each well contained 3 μg protein. (M) Prestained protein marker, ( 1 ) proTHI2.1-TRX, ( 2 ) proTHI2.2-TRX, ( 3 ) proTHI2.3-TRX, ( 4 ) proTHI2.4-TRX
    Figure Legend Snippet: Comparison of Ni–NTA purified proTHI-TRX fusion proteins from strain C3030. a Coomassie Brilliant Blue staining. b Western blot with anti-His tag antibody. Each well contained 3 μg protein. (M) Prestained protein marker, ( 1 ) proTHI2.1-TRX, ( 2 ) proTHI2.2-TRX, ( 3 ) proTHI2.3-TRX, ( 4 ) proTHI2.4-TRX

    Techniques Used: Purification, Staining, Western Blot, Marker

    2) Product Images from "Comparison of periplasmic and intracellular expression of Arabidopsis thionin proproteins in E. coli"

    Article Title: Comparison of periplasmic and intracellular expression of Arabidopsis thionin proproteins in E. coli

    Journal: Biotechnology Letters

    doi: 10.1007/s10529-013-1180-z

    Cytoplamic expression of proTHI-TRX fusion proteins in strain C3030. For all fusion proteins, 1 ml of cell culture was pelleted and dissolved in 100 μl sample buffer. 10 μl from this extract and an equivalent amount for total soluble cytoplasmic fractions were separated on Tricine/SDS gels. (M) Protein marker, ( 1 ) un-induced crude extract, ( 2 ) induced crude extract, ( 3 ) total soluble fraction taken after cell lysis by sonication, ( 4 ) insoluble fraction after sonication. Red stars indicate the 25 kDa fusion protein
    Figure Legend Snippet: Cytoplamic expression of proTHI-TRX fusion proteins in strain C3030. For all fusion proteins, 1 ml of cell culture was pelleted and dissolved in 100 μl sample buffer. 10 μl from this extract and an equivalent amount for total soluble cytoplasmic fractions were separated on Tricine/SDS gels. (M) Protein marker, ( 1 ) un-induced crude extract, ( 2 ) induced crude extract, ( 3 ) total soluble fraction taken after cell lysis by sonication, ( 4 ) insoluble fraction after sonication. Red stars indicate the 25 kDa fusion protein

    Techniques Used: Expressing, Cell Culture, Marker, Lysis, Sonication

    Comparison of Ni–NTA purified proTHI-TRX fusion proteins from strain C3030. a Coomassie Brilliant Blue staining. b Western blot with anti-His tag antibody. Each well contained 3 μg protein. (M) Prestained protein marker, ( 1 ) proTHI2.1-TRX, ( 2 ) proTHI2.2-TRX, ( 3 ) proTHI2.3-TRX, ( 4 ) proTHI2.4-TRX
    Figure Legend Snippet: Comparison of Ni–NTA purified proTHI-TRX fusion proteins from strain C3030. a Coomassie Brilliant Blue staining. b Western blot with anti-His tag antibody. Each well contained 3 μg protein. (M) Prestained protein marker, ( 1 ) proTHI2.1-TRX, ( 2 ) proTHI2.2-TRX, ( 3 ) proTHI2.3-TRX, ( 4 ) proTHI2.4-TRX

    Techniques Used: Purification, Staining, Western Blot, Marker

    Related Articles

    Clone Assay:

    Article Title: Comparison of periplasmic and intracellular expression of Arabidopsis thionin proproteins in E. coli
    Article Snippet: However, E. coli strains with an oxidizing cytoplasm have been reported that result in high levels of functional proteins with disulfide bridges. .. We therefore cloned the expression plasmids for all four fusion proteins into the SHuffle strain C3030 (New England Biolabs) in which the lethal phenotype of lacking the gor and trxB reductases is suppressed by a mutation in the peroxidase ahpC (Faulkner et al. ) thus providing an oxidising environment in the cytoplasm. .. Additionally, expression of periplasmic disulfide bond isomerase (DsbC) in the cytoplasm of these strains is supposed to enhance proper disulfide bond formation (Maskos et al. ).

    Functional Assay:

    Article Title: Comparison of periplasmic and intracellular expression of Arabidopsis thionin proproteins in E. coli
    Article Snippet: However, E. coli strains with an oxidizing cytoplasm have been reported that result in high levels of functional proteins with disulfide bridges. .. We therefore cloned the expression plasmids for all four fusion proteins into the SHuffle strain C3030 (New England Biolabs) in which the lethal phenotype of lacking the gor and trxB reductases is suppressed by a mutation in the peroxidase ahpC (Faulkner et al. ) thus providing an oxidising environment in the cytoplasm.

    Affinity Chromatography:

    Article Title: Comprehensive analysis of Fc-mediated IgM binding to the Plasmodium falciparum erythrocyte membrane protein 1 family in three parasite clones
    Article Snippet: Recombinant proteins with N-terminal his-tags and representing various PfEMP1 domains were produced in Escherichia coli Shuffle C3030 cells (New England Biolabs). .. Constructs were amplified from parasite genomic DNA, using specific primer sets (Table ) and cloned into the pET15b modified plasmid .

    Expressing:

    Article Title: Comparison of periplasmic and intracellular expression of Arabidopsis thionin proproteins in E. coli
    Article Snippet: However, E. coli strains with an oxidizing cytoplasm have been reported that result in high levels of functional proteins with disulfide bridges. .. We therefore cloned the expression plasmids for all four fusion proteins into the SHuffle strain C3030 (New England Biolabs) in which the lethal phenotype of lacking the gor and trxB reductases is suppressed by a mutation in the peroxidase ahpC (Faulkner et al. ) thus providing an oxidising environment in the cytoplasm. .. Additionally, expression of periplasmic disulfide bond isomerase (DsbC) in the cytoplasm of these strains is supposed to enhance proper disulfide bond formation (Maskos et al. ).

    Article Title: Comprehensive analysis of Fc-mediated IgM binding to the Plasmodium falciparum erythrocyte membrane protein 1 family in three parasite clones
    Article Snippet: Paragraph title: Recombinant protein expression ... Recombinant proteins with N-terminal his-tags and representing various PfEMP1 domains were produced in Escherichia coli Shuffle C3030 cells (New England Biolabs).

    Mutagenesis:

    Article Title: Comparison of periplasmic and intracellular expression of Arabidopsis thionin proproteins in E. coli
    Article Snippet: However, E. coli strains with an oxidizing cytoplasm have been reported that result in high levels of functional proteins with disulfide bridges. .. We therefore cloned the expression plasmids for all four fusion proteins into the SHuffle strain C3030 (New England Biolabs) in which the lethal phenotype of lacking the gor and trxB reductases is suppressed by a mutation in the peroxidase ahpC (Faulkner et al. ) thus providing an oxidising environment in the cytoplasm. .. Additionally, expression of periplasmic disulfide bond isomerase (DsbC) in the cytoplasm of these strains is supposed to enhance proper disulfide bond formation (Maskos et al. ).

    Centrifugation:

    Article Title: Comparison of periplasmic and intracellular expression of Arabidopsis thionin proproteins in E. coli
    Article Snippet: We therefore cloned the expression plasmids for all four fusion proteins into the SHuffle strain C3030 (New England Biolabs) in which the lethal phenotype of lacking the gor and trxB reductases is suppressed by a mutation in the peroxidase ahpC (Faulkner et al. ) thus providing an oxidising environment in the cytoplasm. .. Additionally, expression of periplasmic disulfide bond isomerase (DsbC) in the cytoplasm of these strains is supposed to enhance proper disulfide bond formation (Maskos et al. ).

    Labeling:

    Article Title: Comparison of periplasmic and intracellular expression of Arabidopsis thionin proproteins in E. coli
    Article Snippet: These were also labeled with the anti-His-tag antibody and seemed to be degradation products. .. We therefore cloned the expression plasmids for all four fusion proteins into the SHuffle strain C3030 (New England Biolabs) in which the lethal phenotype of lacking the gor and trxB reductases is suppressed by a mutation in the peroxidase ahpC (Faulkner et al. ) thus providing an oxidising environment in the cytoplasm.

    Purification:

    Article Title: Comprehensive analysis of Fc-mediated IgM binding to the Plasmodium falciparum erythrocyte membrane protein 1 family in three parasite clones
    Article Snippet: Recombinant proteins with N-terminal his-tags and representing various PfEMP1 domains were produced in Escherichia coli Shuffle C3030 cells (New England Biolabs). .. Constructs were amplified from parasite genomic DNA, using specific primer sets (Table ) and cloned into the pET15b modified plasmid .

    Article Title: Natural and Vaccine-Induced Acquisition of Cross-Reactive IgG-Inhibiting ICAM-1-Specific Binding of a Plasmodium falciparum PfEMP1 Subtype Associated Specifically with Cerebral Malaria
    Article Snippet: Amplicons were subcloned into a modified pET15b vector and expressed as His-tagged proteins in Escherichia coli Shuffle C3030 cells (New England BioLabs) as described previously ( ). .. All the proteins were purified (see Fig. S1 in the supplemental material) by immobilized metal ion affinity chromatography using HisTrap HP 1-ml columns (GE Healthcare) and are referred to by the designations listed in .

    Produced:

    Article Title: Comparison of periplasmic and intracellular expression of Arabidopsis thionin proproteins in E. coli
    Article Snippet: However, the fraction of soluble fusion protein was larger than that of the Rosetta strain (Supplementary Fig. 5). .. The amount of fusion protein that could be produced from the SHuffle strain C3030 is shown in Table and was higher than obtained with Rosetta(DE3)pLysS. .. Western blotting of the purified fusion proteins (Fig. ) showed less contaminating proteins than with the Rosetta(DE3)pLysS strain (Supplementary Fig. 6).

    Article Title: Comprehensive analysis of Fc-mediated IgM binding to the Plasmodium falciparum erythrocyte membrane protein 1 family in three parasite clones
    Article Snippet: The transcription levels for each var gene was calculated using the 2−ΔCT method, relative to the levels of the housekeeping gene seryl-tRNA synthetase. .. Recombinant proteins with N-terminal his-tags and representing various PfEMP1 domains were produced in Escherichia coli Shuffle C3030 cells (New England Biolabs). .. Constructs were amplified from parasite genomic DNA, using specific primer sets (Table ) and cloned into the pET15b modified plasmid .

    Article Title: Natural and Vaccine-Induced Acquisition of Cross-Reactive IgG-Inhibiting ICAM-1-Specific Binding of a Plasmodium falciparum PfEMP1 Subtype Associated Specifically with Cerebral Malaria
    Article Snippet: The genes encoding the DBLβ domains used were amplified from genomic DNA or produced as synthetic genes ( ) ( ). .. Amplicons were subcloned into a modified pET15b vector and expressed as His-tagged proteins in Escherichia coli Shuffle C3030 cells (New England BioLabs) as described previously ( ).

    Amplification:

    Article Title: Natural and Vaccine-Induced Acquisition of Cross-Reactive IgG-Inhibiting ICAM-1-Specific Binding of a Plasmodium falciparum PfEMP1 Subtype Associated Specifically with Cerebral Malaria
    Article Snippet: The genes encoding the DBLβ domains used were amplified from genomic DNA or produced as synthetic genes ( ) ( ). .. Amplicons were subcloned into a modified pET15b vector and expressed as His-tagged proteins in Escherichia coli Shuffle C3030 cells (New England BioLabs) as described previously ( ).

    Polyacrylamide Gel Electrophoresis:

    Article Title: Comprehensive analysis of Fc-mediated IgM binding to the Plasmodium falciparum erythrocyte membrane protein 1 family in three parasite clones
    Article Snippet: Recombinant proteins with N-terminal his-tags and representing various PfEMP1 domains were produced in Escherichia coli Shuffle C3030 cells (New England Biolabs). .. All expressed proteins were purified by immobilized metal ion affinity chromatography using His-Trap High Performance columns (GE Healthcare).

    Western Blot:

    Article Title: Comparison of periplasmic and intracellular expression of Arabidopsis thionin proproteins in E. coli
    Article Snippet: Western blotting with the anti-His-tag antibody also detected a large band in the proTHI2.1 fraction at around 60 kDa and a smaller (20 kDa) and larger band (30 kDa) in the proTHI2.3 fraction. .. We therefore cloned the expression plasmids for all four fusion proteins into the SHuffle strain C3030 (New England Biolabs) in which the lethal phenotype of lacking the gor and trxB reductases is suppressed by a mutation in the peroxidase ahpC (Faulkner et al. ) thus providing an oxidising environment in the cytoplasm.

    Article Title: Comprehensive analysis of Fc-mediated IgM binding to the Plasmodium falciparum erythrocyte membrane protein 1 family in three parasite clones
    Article Snippet: Recombinant proteins with N-terminal his-tags and representing various PfEMP1 domains were produced in Escherichia coli Shuffle C3030 cells (New England Biolabs). .. The purity of each protein was analyzed by sodium-dodecyl-sulfate polyacrylamide gel electrophoresis, followed by Coomassie staining with InstantBlue (Expedeon) following the manufacturer’s instructions.

    Modification:

    Article Title: Natural and Vaccine-Induced Acquisition of Cross-Reactive IgG-Inhibiting ICAM-1-Specific Binding of a Plasmodium falciparum PfEMP1 Subtype Associated Specifically with Cerebral Malaria
    Article Snippet: The genes encoding the DBLβ domains used were amplified from genomic DNA or produced as synthetic genes ( ) ( ). .. Amplicons were subcloned into a modified pET15b vector and expressed as His-tagged proteins in Escherichia coli Shuffle C3030 cells (New England BioLabs) as described previously ( ). .. All the proteins were purified (see Fig. S1 in the supplemental material) by immobilized metal ion affinity chromatography using HisTrap HP 1-ml columns (GE Healthcare) and are referred to by the designations listed in .

    Staining:

    Article Title: Comprehensive analysis of Fc-mediated IgM binding to the Plasmodium falciparum erythrocyte membrane protein 1 family in three parasite clones
    Article Snippet: Recombinant proteins with N-terminal his-tags and representing various PfEMP1 domains were produced in Escherichia coli Shuffle C3030 cells (New England Biolabs). .. All expressed proteins were purified by immobilized metal ion affinity chromatography using His-Trap High Performance columns (GE Healthcare).

    Recombinant:

    Article Title: Comprehensive analysis of Fc-mediated IgM binding to the Plasmodium falciparum erythrocyte membrane protein 1 family in three parasite clones
    Article Snippet: The transcription levels for each var gene was calculated using the 2−ΔCT method, relative to the levels of the housekeeping gene seryl-tRNA synthetase. .. Recombinant proteins with N-terminal his-tags and representing various PfEMP1 domains were produced in Escherichia coli Shuffle C3030 cells (New England Biolabs). .. Constructs were amplified from parasite genomic DNA, using specific primer sets (Table ) and cloned into the pET15b modified plasmid .

    Article Title: Natural and Vaccine-Induced Acquisition of Cross-Reactive IgG-Inhibiting ICAM-1-Specific Binding of a Plasmodium falciparum PfEMP1 Subtype Associated Specifically with Cerebral Malaria
    Article Snippet: Paragraph title: Recombinant proteins. ... Amplicons were subcloned into a modified pET15b vector and expressed as His-tagged proteins in Escherichia coli Shuffle C3030 cells (New England BioLabs) as described previously ( ).

    Plasmid Preparation:

    Article Title: Natural and Vaccine-Induced Acquisition of Cross-Reactive IgG-Inhibiting ICAM-1-Specific Binding of a Plasmodium falciparum PfEMP1 Subtype Associated Specifically with Cerebral Malaria
    Article Snippet: The genes encoding the DBLβ domains used were amplified from genomic DNA or produced as synthetic genes ( ) ( ). .. Amplicons were subcloned into a modified pET15b vector and expressed as His-tagged proteins in Escherichia coli Shuffle C3030 cells (New England BioLabs) as described previously ( ). .. All the proteins were purified (see Fig. S1 in the supplemental material) by immobilized metal ion affinity chromatography using HisTrap HP 1-ml columns (GE Healthcare) and are referred to by the designations listed in .

    Flow Cytometry:

    Article Title: Natural and Vaccine-Induced Acquisition of Cross-Reactive IgG-Inhibiting ICAM-1-Specific Binding of a Plasmodium falciparum PfEMP1 Subtype Associated Specifically with Cerebral Malaria
    Article Snippet: Amplicons were subcloned into a modified pET15b vector and expressed as His-tagged proteins in Escherichia coli Shuffle C3030 cells (New England BioLabs) as described previously ( ). .. All the proteins were purified (see Fig. S1 in the supplemental material) by immobilized metal ion affinity chromatography using HisTrap HP 1-ml columns (GE Healthcare) and are referred to by the designations listed in .

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    New England Biolabs escherichia coli shuffle c3030 cells
    Escherichia Coli Shuffle C3030 Cells, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/escherichia coli shuffle c3030 cells/product/New England Biolabs
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    New England Biolabs shuffle strain c3030
    Cytoplamic expression of proTHI-TRX fusion proteins in strain <t>C3030.</t> For all fusion proteins, 1 ml of cell culture was pelleted and dissolved in 100 μl sample buffer. 10 μl from this extract and an equivalent amount for total soluble cytoplasmic fractions were separated on Tricine/SDS gels. (M) Protein marker, ( 1 ) un-induced crude extract, ( 2 ) induced crude extract, ( 3 ) total soluble fraction taken after cell lysis by sonication, ( 4 ) insoluble fraction after sonication. Red stars indicate the 25 kDa fusion protein
    Shuffle Strain C3030, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 78/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/shuffle strain c3030/product/New England Biolabs
    Average 78 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    shuffle strain c3030 - by Bioz Stars, 2019-07
    78/100 stars
      Buy from Supplier

    Image Search Results


    Cytoplamic expression of proTHI-TRX fusion proteins in strain C3030. For all fusion proteins, 1 ml of cell culture was pelleted and dissolved in 100 μl sample buffer. 10 μl from this extract and an equivalent amount for total soluble cytoplasmic fractions were separated on Tricine/SDS gels. (M) Protein marker, ( 1 ) un-induced crude extract, ( 2 ) induced crude extract, ( 3 ) total soluble fraction taken after cell lysis by sonication, ( 4 ) insoluble fraction after sonication. Red stars indicate the 25 kDa fusion protein

    Journal: Biotechnology Letters

    Article Title: Comparison of periplasmic and intracellular expression of Arabidopsis thionin proproteins in E. coli

    doi: 10.1007/s10529-013-1180-z

    Figure Lengend Snippet: Cytoplamic expression of proTHI-TRX fusion proteins in strain C3030. For all fusion proteins, 1 ml of cell culture was pelleted and dissolved in 100 μl sample buffer. 10 μl from this extract and an equivalent amount for total soluble cytoplasmic fractions were separated on Tricine/SDS gels. (M) Protein marker, ( 1 ) un-induced crude extract, ( 2 ) induced crude extract, ( 3 ) total soluble fraction taken after cell lysis by sonication, ( 4 ) insoluble fraction after sonication. Red stars indicate the 25 kDa fusion protein

    Article Snippet: We therefore cloned the expression plasmids for all four fusion proteins into the SHuffle strain C3030 (New England Biolabs) in which the lethal phenotype of lacking the gor and trxB reductases is suppressed by a mutation in the peroxidase ahpC (Faulkner et al. ) thus providing an oxidising environment in the cytoplasm.

    Techniques: Expressing, Cell Culture, Marker, Lysis, Sonication

    Comparison of Ni–NTA purified proTHI-TRX fusion proteins from strain C3030. a Coomassie Brilliant Blue staining. b Western blot with anti-His tag antibody. Each well contained 3 μg protein. (M) Prestained protein marker, ( 1 ) proTHI2.1-TRX, ( 2 ) proTHI2.2-TRX, ( 3 ) proTHI2.3-TRX, ( 4 ) proTHI2.4-TRX

    Journal: Biotechnology Letters

    Article Title: Comparison of periplasmic and intracellular expression of Arabidopsis thionin proproteins in E. coli

    doi: 10.1007/s10529-013-1180-z

    Figure Lengend Snippet: Comparison of Ni–NTA purified proTHI-TRX fusion proteins from strain C3030. a Coomassie Brilliant Blue staining. b Western blot with anti-His tag antibody. Each well contained 3 μg protein. (M) Prestained protein marker, ( 1 ) proTHI2.1-TRX, ( 2 ) proTHI2.2-TRX, ( 3 ) proTHI2.3-TRX, ( 4 ) proTHI2.4-TRX

    Article Snippet: We therefore cloned the expression plasmids for all four fusion proteins into the SHuffle strain C3030 (New England Biolabs) in which the lethal phenotype of lacking the gor and trxB reductases is suppressed by a mutation in the peroxidase ahpC (Faulkner et al. ) thus providing an oxidising environment in the cytoplasm.

    Techniques: Purification, Staining, Western Blot, Marker