shuffle e coli strain  (New England Biolabs)


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    Name:
    SHuffle Express Competent E coli
    Description:
    SHuffle Express Competent E coli 12x0 05 ml
    Catalog Number:
    c3028j
    Price:
    228
    Size:
    12x0 05 ml
    Category:
    Competent Bacteria
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    Structured Review

    New England Biolabs shuffle e coli strain
    SHuffle Express Competent E coli
    SHuffle Express Competent E coli 12x0 05 ml
    https://www.bioz.com/result/shuffle e coli strain/product/New England Biolabs
    Average 97 stars, based on 119 article reviews
    Price from $9.99 to $1999.99
    shuffle e coli strain - by Bioz Stars, 2020-09
    97/100 stars

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    1) Product Images from "Soluble expression of recombinant midgut zymogen (native propeptide) proteases from the Aedes aegypti Mosquito Utilizing E. coli as a host"

    Article Title: Soluble expression of recombinant midgut zymogen (native propeptide) proteases from the Aedes aegypti Mosquito Utilizing E. coli as a host

    Journal: BMC Biochemistry

    doi: 10.1186/s12858-018-0101-0

    SDS-PAGE analysis and BApNA activity assays of samples collected from small-scale growth experiments of SHuffle®  E. coli  cells (NEB) grown in TB media at 15 °C (induced with 0.1 mM IPTG). Samples were collected at the given time points (in hours). The MW ladder is in kilo-Daltons (kDa).  a  The gel represents the soluble expression of AaET-NL zymogen (MW ~ 27.0 kDa, yellow arrow), auto-activating to the active mature form (MW ~ 22.4 kDa, purple arrow). The presence of active AaET at 5 h post-induction correlates with an increase in BApNA activity, with maximal activity observed at the 24 h time-point (plot on the right).  b  The gel represents the soluble expression of AaSPVI-NL zymogen (MW ~ 28.7 kDa, yellow arrow), auto-activating to the active mature form (MW ~ 24.1 kDa, purple arrow). The presence of active AaSPVI at 16 h post-induction correlates with an increase in BApNA activity, with maximal activity observed at the 67 h time-point (plot on the right).  c  The gel represents the soluble expression of AaSPVII-NL zymogen (MW ~ 28.7 kDa, yellow arrow), auto-activating to the active mature form (MW ~ 24.2 kDa, purple arrow). The presence of active AaSPVII at 15 h post-induction correlates with an increase in BApNA activity, with maximal activity observed at the 18 h time-point (plot on the right). Unlike AaET and AaSPVI, AaSPVII expression results in a species that lies between the zymogen and active forms starting at 8 h post-induction and disappearing at 15 h. This species is an inactive form of AaSPVII since no detectable BApNA activity observed
    Figure Legend Snippet: SDS-PAGE analysis and BApNA activity assays of samples collected from small-scale growth experiments of SHuffle® E. coli cells (NEB) grown in TB media at 15 °C (induced with 0.1 mM IPTG). Samples were collected at the given time points (in hours). The MW ladder is in kilo-Daltons (kDa). a The gel represents the soluble expression of AaET-NL zymogen (MW ~ 27.0 kDa, yellow arrow), auto-activating to the active mature form (MW ~ 22.4 kDa, purple arrow). The presence of active AaET at 5 h post-induction correlates with an increase in BApNA activity, with maximal activity observed at the 24 h time-point (plot on the right). b The gel represents the soluble expression of AaSPVI-NL zymogen (MW ~ 28.7 kDa, yellow arrow), auto-activating to the active mature form (MW ~ 24.1 kDa, purple arrow). The presence of active AaSPVI at 16 h post-induction correlates with an increase in BApNA activity, with maximal activity observed at the 67 h time-point (plot on the right). c The gel represents the soluble expression of AaSPVII-NL zymogen (MW ~ 28.7 kDa, yellow arrow), auto-activating to the active mature form (MW ~ 24.2 kDa, purple arrow). The presence of active AaSPVII at 15 h post-induction correlates with an increase in BApNA activity, with maximal activity observed at the 18 h time-point (plot on the right). Unlike AaET and AaSPVI, AaSPVII expression results in a species that lies between the zymogen and active forms starting at 8 h post-induction and disappearing at 15 h. This species is an inactive form of AaSPVII since no detectable BApNA activity observed

    Techniques Used: SDS Page, Activity Assay, Expressing

    Related Articles

    Clone Assay:

    Article Title: Soluble expression of recombinant midgut zymogen (native propeptide) proteases from the Aedes aegypti Mosquito Utilizing E. coli as a host
    Article Snippet: .. Expression of recombinant proteases using SHuffle® (NEB) E. coli cells and Bis-Tris gel analysis The most commonly used bacterial strain for expression of genes cloned into the pET vector system are the BL21(DE3) and E. coli K12 lineage strains [ ]. ..

    Positron Emission Tomography:

    Article Title: Soluble expression of recombinant midgut zymogen (native propeptide) proteases from the Aedes aegypti Mosquito Utilizing E. coli as a host
    Article Snippet: .. Expression of recombinant proteases using SHuffle® (NEB) E. coli cells and Bis-Tris gel analysis The most commonly used bacterial strain for expression of genes cloned into the pET vector system are the BL21(DE3) and E. coli K12 lineage strains [ ]. ..

    Mutagenesis:

    Article Title: A Dual-Intein Autoprocessing Domain that Directs Synchronized Protein Co-Expression in Both Prokaryotes and Eukaryotes
    Article Snippet: .. In addition to E. coli BL21(DE3), protein expression was performed in SHuffle E. coli strain (New England Biolab, Ipswich, MA) lacking the trxB and gor reductases along with an additional suppressor mutation (ahpC) which restores viability . ..

    Isolation:

    Article Title: Development of an Antigen Detection ELISA for Bancroftian Filariasis Using BmSXP-Specific Recombinant Monoclonal Antibody
    Article Snippet: .. The 5B recombinant monoclonal antibody protein was isolated using phage display technology and produced as previously described., Briefly, the 5B monoclonal plasmid was transformed into SHuffle® Express Escherichia coli cells (New England Biolabs) and expression was performed in 1 L 2-YT broth supplemented with 100 μg/mL ampicillin and 0.2% glucose. .. The culture was grown at 37°C at 200 rpm until the OD600 reached 0.6–0.7, then induced with 1 mM isopropyl-B-D-thiogalactopyranoside, and further cultured for 16 hours at 25°C with 180 rpm shaking.

    Purification:

    Article Title: Nanobodies: site-specific labeling for super-resolution imaging, rapid epitope-mapping and native protein complex isolation
    Article Snippet: .. Expression and purification of nanobodies Nanobodies with protease-cleavable affinity tags or engineered cysteines were routinely expressed in the cytoplasm of E. coli BLR (BL21 derivative; Novagen) or E. coli SHuffle Express (New England Biolabs). ..

    Produced:

    Article Title: Development of an Antigen Detection ELISA for Bancroftian Filariasis Using BmSXP-Specific Recombinant Monoclonal Antibody
    Article Snippet: .. The 5B recombinant monoclonal antibody protein was isolated using phage display technology and produced as previously described., Briefly, the 5B monoclonal plasmid was transformed into SHuffle® Express Escherichia coli cells (New England Biolabs) and expression was performed in 1 L 2-YT broth supplemented with 100 μg/mL ampicillin and 0.2% glucose. .. The culture was grown at 37°C at 200 rpm until the OD600 reached 0.6–0.7, then induced with 1 mM isopropyl-B-D-thiogalactopyranoside, and further cultured for 16 hours at 25°C with 180 rpm shaking.

    Inhibition:

    Article Title: The rare sugar d-tagatose protects plants from downy mildews and is a safe fungicidal agrochemical
    Article Snippet: .. Characterizations of enzymes in d -fructose 6-phosphate metabolic pathway and inhibition of activity by d -tagatose 6-phosphate The cDNA fragment (described above) of the coding region from phosphomannose isomerase (LC500563) of H. arabidopsidis isolate Noco2 was subcloned in frame into the pColdI vector (Takara), and expressed in E. coli SHuffle Express Competent cells (New England BioLabs, MA, USA) according to the manufacturer’s instructions. .. The recombinant proteins of the coding region generated by the heterologous expression system were obtained using a mass culture system with 10 liters LB broth containing 1% (w/v) d -glucose and 100 µg ml−1 carbenicillin at 30 °C and 200 rpm for 6 h (preculture), 15 °C and 200 rpm for 1 h (cold shock induction), and 15 °C and 200 rpm for 3 days in Jar Fermenter TS-M15L and purified using a HisTrap HP column (GE Healthcare) as per the manufacturer’s instructions, then dialyzed against 10 mM Tris-HCl buffer (pH 7.4).

    Activity Assay:

    Article Title: The rare sugar d-tagatose protects plants from downy mildews and is a safe fungicidal agrochemical
    Article Snippet: .. Characterizations of enzymes in d -fructose 6-phosphate metabolic pathway and inhibition of activity by d -tagatose 6-phosphate The cDNA fragment (described above) of the coding region from phosphomannose isomerase (LC500563) of H. arabidopsidis isolate Noco2 was subcloned in frame into the pColdI vector (Takara), and expressed in E. coli SHuffle Express Competent cells (New England BioLabs, MA, USA) according to the manufacturer’s instructions. .. The recombinant proteins of the coding region generated by the heterologous expression system were obtained using a mass culture system with 10 liters LB broth containing 1% (w/v) d -glucose and 100 µg ml−1 carbenicillin at 30 °C and 200 rpm for 6 h (preculture), 15 °C and 200 rpm for 1 h (cold shock induction), and 15 °C and 200 rpm for 3 days in Jar Fermenter TS-M15L and purified using a HisTrap HP column (GE Healthcare) as per the manufacturer’s instructions, then dialyzed against 10 mM Tris-HCl buffer (pH 7.4).

    Expressing:

    Article Title: Structures of Xenopus Embryonic Epidermal Lectin Reveal a Conserved Mechanism of Microbial Glycan Recognition *
    Article Snippet: .. The expression plasmid was transformed into Escherichia coli SHuffle® Express (New England Biolabs). ..

    Article Title: Soluble expression of recombinant midgut zymogen (native propeptide) proteases from the Aedes aegypti Mosquito Utilizing E. coli as a host
    Article Snippet: .. Expression of recombinant proteases using SHuffle® (NEB) E. coli cells and Bis-Tris gel analysis The most commonly used bacterial strain for expression of genes cloned into the pET vector system are the BL21(DE3) and E. coli K12 lineage strains [ ]. ..

    Article Title: A Dual-Intein Autoprocessing Domain that Directs Synchronized Protein Co-Expression in Both Prokaryotes and Eukaryotes
    Article Snippet: .. In addition to E. coli BL21(DE3), protein expression was performed in SHuffle E. coli strain (New England Biolab, Ipswich, MA) lacking the trxB and gor reductases along with an additional suppressor mutation (ahpC) which restores viability . ..

    Article Title: Nanobodies: site-specific labeling for super-resolution imaging, rapid epitope-mapping and native protein complex isolation
    Article Snippet: .. Expression and purification of nanobodies Nanobodies with protease-cleavable affinity tags or engineered cysteines were routinely expressed in the cytoplasm of E. coli BLR (BL21 derivative; Novagen) or E. coli SHuffle Express (New England Biolabs). ..

    Article Title: Development of an Antigen Detection ELISA for Bancroftian Filariasis Using BmSXP-Specific Recombinant Monoclonal Antibody
    Article Snippet: .. The 5B recombinant monoclonal antibody protein was isolated using phage display technology and produced as previously described., Briefly, the 5B monoclonal plasmid was transformed into SHuffle® Express Escherichia coli cells (New England Biolabs) and expression was performed in 1 L 2-YT broth supplemented with 100 μg/mL ampicillin and 0.2% glucose. .. The culture was grown at 37°C at 200 rpm until the OD600 reached 0.6–0.7, then induced with 1 mM isopropyl-B-D-thiogalactopyranoside, and further cultured for 16 hours at 25°C with 180 rpm shaking.

    Transformation Assay:

    Article Title: Structures of Xenopus Embryonic Epidermal Lectin Reveal a Conserved Mechanism of Microbial Glycan Recognition *
    Article Snippet: .. The expression plasmid was transformed into Escherichia coli SHuffle® Express (New England Biolabs). ..

    Article Title: Development of an Antigen Detection ELISA for Bancroftian Filariasis Using BmSXP-Specific Recombinant Monoclonal Antibody
    Article Snippet: .. The 5B recombinant monoclonal antibody protein was isolated using phage display technology and produced as previously described., Briefly, the 5B monoclonal plasmid was transformed into SHuffle® Express Escherichia coli cells (New England Biolabs) and expression was performed in 1 L 2-YT broth supplemented with 100 μg/mL ampicillin and 0.2% glucose. .. The culture was grown at 37°C at 200 rpm until the OD600 reached 0.6–0.7, then induced with 1 mM isopropyl-B-D-thiogalactopyranoside, and further cultured for 16 hours at 25°C with 180 rpm shaking.

    Recombinant:

    Article Title: Soluble expression of recombinant midgut zymogen (native propeptide) proteases from the Aedes aegypti Mosquito Utilizing E. coli as a host
    Article Snippet: .. Expression of recombinant proteases using SHuffle® (NEB) E. coli cells and Bis-Tris gel analysis The most commonly used bacterial strain for expression of genes cloned into the pET vector system are the BL21(DE3) and E. coli K12 lineage strains [ ]. ..

    Article Title: Development of an Antigen Detection ELISA for Bancroftian Filariasis Using BmSXP-Specific Recombinant Monoclonal Antibody
    Article Snippet: .. The 5B recombinant monoclonal antibody protein was isolated using phage display technology and produced as previously described., Briefly, the 5B monoclonal plasmid was transformed into SHuffle® Express Escherichia coli cells (New England Biolabs) and expression was performed in 1 L 2-YT broth supplemented with 100 μg/mL ampicillin and 0.2% glucose. .. The culture was grown at 37°C at 200 rpm until the OD600 reached 0.6–0.7, then induced with 1 mM isopropyl-B-D-thiogalactopyranoside, and further cultured for 16 hours at 25°C with 180 rpm shaking.

    Plasmid Preparation:

    Article Title: Structures of Xenopus Embryonic Epidermal Lectin Reveal a Conserved Mechanism of Microbial Glycan Recognition *
    Article Snippet: .. The expression plasmid was transformed into Escherichia coli SHuffle® Express (New England Biolabs). ..

    Article Title: Soluble expression of recombinant midgut zymogen (native propeptide) proteases from the Aedes aegypti Mosquito Utilizing E. coli as a host
    Article Snippet: .. Expression of recombinant proteases using SHuffle® (NEB) E. coli cells and Bis-Tris gel analysis The most commonly used bacterial strain for expression of genes cloned into the pET vector system are the BL21(DE3) and E. coli K12 lineage strains [ ]. ..

    Article Title: The rare sugar d-tagatose protects plants from downy mildews and is a safe fungicidal agrochemical
    Article Snippet: .. Characterizations of enzymes in d -fructose 6-phosphate metabolic pathway and inhibition of activity by d -tagatose 6-phosphate The cDNA fragment (described above) of the coding region from phosphomannose isomerase (LC500563) of H. arabidopsidis isolate Noco2 was subcloned in frame into the pColdI vector (Takara), and expressed in E. coli SHuffle Express Competent cells (New England BioLabs, MA, USA) according to the manufacturer’s instructions. .. The recombinant proteins of the coding region generated by the heterologous expression system were obtained using a mass culture system with 10 liters LB broth containing 1% (w/v) d -glucose and 100 µg ml−1 carbenicillin at 30 °C and 200 rpm for 6 h (preculture), 15 °C and 200 rpm for 1 h (cold shock induction), and 15 °C and 200 rpm for 3 days in Jar Fermenter TS-M15L and purified using a HisTrap HP column (GE Healthcare) as per the manufacturer’s instructions, then dialyzed against 10 mM Tris-HCl buffer (pH 7.4).

    Article Title: Development of an Antigen Detection ELISA for Bancroftian Filariasis Using BmSXP-Specific Recombinant Monoclonal Antibody
    Article Snippet: .. The 5B recombinant monoclonal antibody protein was isolated using phage display technology and produced as previously described., Briefly, the 5B monoclonal plasmid was transformed into SHuffle® Express Escherichia coli cells (New England Biolabs) and expression was performed in 1 L 2-YT broth supplemented with 100 μg/mL ampicillin and 0.2% glucose. .. The culture was grown at 37°C at 200 rpm until the OD600 reached 0.6–0.7, then induced with 1 mM isopropyl-B-D-thiogalactopyranoside, and further cultured for 16 hours at 25°C with 180 rpm shaking.

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    New England Biolabs shuffle e coli strain
    SDS-PAGE analysis and BApNA activity assays of samples collected from small-scale growth experiments of SHuffle®  E. coli  cells (NEB) grown in TB media at 15 °C (induced with 0.1 mM IPTG). Samples were collected at the given time points (in hours). The MW ladder is in kilo-Daltons (kDa).  a  The gel represents the soluble expression of AaET-NL zymogen (MW ~ 27.0 kDa, yellow arrow), auto-activating to the active mature form (MW ~ 22.4 kDa, purple arrow). The presence of active AaET at 5 h post-induction correlates with an increase in BApNA activity, with maximal activity observed at the 24 h time-point (plot on the right).  b  The gel represents the soluble expression of AaSPVI-NL zymogen (MW ~ 28.7 kDa, yellow arrow), auto-activating to the active mature form (MW ~ 24.1 kDa, purple arrow). The presence of active AaSPVI at 16 h post-induction correlates with an increase in BApNA activity, with maximal activity observed at the 67 h time-point (plot on the right).  c  The gel represents the soluble expression of AaSPVII-NL zymogen (MW ~ 28.7 kDa, yellow arrow), auto-activating to the active mature form (MW ~ 24.2 kDa, purple arrow). The presence of active AaSPVII at 15 h post-induction correlates with an increase in BApNA activity, with maximal activity observed at the 18 h time-point (plot on the right). Unlike AaET and AaSPVI, AaSPVII expression results in a species that lies between the zymogen and active forms starting at 8 h post-induction and disappearing at 15 h. This species is an inactive form of AaSPVII since no detectable BApNA activity observed
    Shuffle E Coli Strain, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 97/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/shuffle e coli strain/product/New England Biolabs
    Average 97 stars, based on 14 article reviews
    Price from $9.99 to $1999.99
    shuffle e coli strain - by Bioz Stars, 2020-09
    97/100 stars
      Buy from Supplier

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    SDS-PAGE analysis and BApNA activity assays of samples collected from small-scale growth experiments of SHuffle®  E. coli  cells (NEB) grown in TB media at 15 °C (induced with 0.1 mM IPTG). Samples were collected at the given time points (in hours). The MW ladder is in kilo-Daltons (kDa).  a  The gel represents the soluble expression of AaET-NL zymogen (MW ~ 27.0 kDa, yellow arrow), auto-activating to the active mature form (MW ~ 22.4 kDa, purple arrow). The presence of active AaET at 5 h post-induction correlates with an increase in BApNA activity, with maximal activity observed at the 24 h time-point (plot on the right).  b  The gel represents the soluble expression of AaSPVI-NL zymogen (MW ~ 28.7 kDa, yellow arrow), auto-activating to the active mature form (MW ~ 24.1 kDa, purple arrow). The presence of active AaSPVI at 16 h post-induction correlates with an increase in BApNA activity, with maximal activity observed at the 67 h time-point (plot on the right).  c  The gel represents the soluble expression of AaSPVII-NL zymogen (MW ~ 28.7 kDa, yellow arrow), auto-activating to the active mature form (MW ~ 24.2 kDa, purple arrow). The presence of active AaSPVII at 15 h post-induction correlates with an increase in BApNA activity, with maximal activity observed at the 18 h time-point (plot on the right). Unlike AaET and AaSPVI, AaSPVII expression results in a species that lies between the zymogen and active forms starting at 8 h post-induction and disappearing at 15 h. This species is an inactive form of AaSPVII since no detectable BApNA activity observed

    Journal: BMC Biochemistry

    Article Title: Soluble expression of recombinant midgut zymogen (native propeptide) proteases from the Aedes aegypti Mosquito Utilizing E. coli as a host

    doi: 10.1186/s12858-018-0101-0

    Figure Lengend Snippet: SDS-PAGE analysis and BApNA activity assays of samples collected from small-scale growth experiments of SHuffle® E. coli cells (NEB) grown in TB media at 15 °C (induced with 0.1 mM IPTG). Samples were collected at the given time points (in hours). The MW ladder is in kilo-Daltons (kDa). a The gel represents the soluble expression of AaET-NL zymogen (MW ~ 27.0 kDa, yellow arrow), auto-activating to the active mature form (MW ~ 22.4 kDa, purple arrow). The presence of active AaET at 5 h post-induction correlates with an increase in BApNA activity, with maximal activity observed at the 24 h time-point (plot on the right). b The gel represents the soluble expression of AaSPVI-NL zymogen (MW ~ 28.7 kDa, yellow arrow), auto-activating to the active mature form (MW ~ 24.1 kDa, purple arrow). The presence of active AaSPVI at 16 h post-induction correlates with an increase in BApNA activity, with maximal activity observed at the 67 h time-point (plot on the right). c The gel represents the soluble expression of AaSPVII-NL zymogen (MW ~ 28.7 kDa, yellow arrow), auto-activating to the active mature form (MW ~ 24.2 kDa, purple arrow). The presence of active AaSPVII at 15 h post-induction correlates with an increase in BApNA activity, with maximal activity observed at the 18 h time-point (plot on the right). Unlike AaET and AaSPVI, AaSPVII expression results in a species that lies between the zymogen and active forms starting at 8 h post-induction and disappearing at 15 h. This species is an inactive form of AaSPVII since no detectable BApNA activity observed

    Article Snippet: With engineering of the SHuffle E. coli strain (New England Biolabs), with a more oxidizing cytoplasm, the four most abundant proteases were successfully solubly expressed in a bacterial system.

    Techniques: SDS Page, Activity Assay, Expressing