bl21 de3 bacteria  (Millipore)


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    Structured Review

    Millipore bl21 de3 bacteria
    Bl21 De3 Bacteria, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bl21 de3 bacteria/product/Millipore
    Average 92 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    bl21 de3 bacteria - by Bioz Stars, 2020-04
    92/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Characterization of ubiquitin and ubiquitin-like-protein isopeptidase activities
    Article Snippet: .. The mature mouse group X PLA2 (amino acids 29–151) was cloned and expressed in BL21 (DE3) bacteria (Novagen) using standard molecular biology techniques (see Supplemental Table 2 for primer information). .. 6His-Ub-PLA2 was expressed and purified as described ( ; ).

    Article Title: EarlyCDT(R)-Lung test: improved clinical utility through additional autoantibody assays
    Article Snippet: The proteins p53, NY-ESO-1, CAGE, Annexin I, MAGE A4, HuD, SOX2-B, and GBU4-5 were cloned into pET21b and produced with a His tag and BirA tag [ , ], whereas SOX2-N was cloned into pET44b and produced with a His tag and NusA tag. .. The recombinant proteins were expressed in BL21 (DE3) bacteria (Novagen) and grown in terrific broth (TB), autoinduction TB media (Novagen), ECPM media, or Power Broth (Molecular Dimensions).

    Centrifugation:

    Article Title: The T-LAK Cell-originated Protein Kinase Signal Pathway Promotes Colorectal Cancer Metastasis
    Article Snippet: 2.5 Bacterial Expression and Purification of Fusion Proteins His-tagged proteins, including pQE81L-PRPK, pET46-WtPRPK, pET46-MutPRPK (S250A), were expressed in BL21 (DE3) bacteria (Novagen, USA) and purified as described for purification of peroxiredoxin 1 (Prx1) ( ). .. Bacteria were grown at 37 °C to an absorbance of 0.9–1.0 at 600 nm and induced with 0.5 mM IPTG overnight at 15 °C and harvested by centrifugation.

    Article Title: The T300A Crohn's disease risk polymorphism impairs function of the WD40 domain of ATG16L1
    Article Snippet: Lysates were cleared by centrifugation and incubated (1 h, 4 °C, rotation) with agarose beads coupled to glutathione (GE Healthcare). .. Loaded beads were washed extensively and used for ATG16L1 pull-down from induced crude bacterial lysates containing ATG16L1-T300 or -A300 proteins expressed from the pET plasmid (Novagen) in BL21(DE3) bacteria.

    Article Title: T-LAK Cell-originated Protein Kinase (TOPK) Phosphorylation of Prx1 at Ser-32 Prevents UVB-induced Apoptosis in RPMI7951 Melanoma Cells through the Regulation of Prx1 Peroxidase Activity *
    Article Snippet: His-Prx1-Wt (Wt), His-Prx1-S32A (S32A), His Prx1-S126A (S126A), His-Prx3 fusion proteins or His-Prx1-deletion mutant proteins (Ndel , Cdel or Δdel ) were expressed in BL21 (DE3) bacteria (Novagen). .. Bacteria were grown at 37 °C to an absorbance of 0.9–1.0 at 600 nm and induced with 0.5 m m IPTG overnight at 15 °C and harvested by centrifugation.

    Amplification:

    Article Title: NC1 domain of collagen ?3(IV) derived from the basement membrane regulates Sertoli cell blood-testis barrier dynamics
    Article Snippet: Production of antibody against Colα3(IV) NC1 domain Recombinant Colα3(IV) NC1 domain was produced using the pET30 Ek/LIC vector kit in BL21 (DE3) bacteria (EMD Millipore). .. Colα3(IV) NC1 domain was amplified by sense 5′-GACGACGACAAGATCACAAGAATGAGAGGCTTCATCTTCA-3′ and antisense 5′-GAGGAGAAGCCCGGTGTGTCTTTTCTTCATGCACACCTGA-3′ primers using 90-d-old rat testis cDNA as template.

    Binding Assay:

    Article Title: Nutrient Isothiocyanates Covalently Modify and Inhibit the Inflammatory Cytokine Macrophage Migration Inhibitory Factor (MIF)
    Article Snippet: In some cases, the MIF coding region was fused to a chitin binding domain (CBD) tag at the C terminus for purification on chitin beads. .. Recombinant MIF with a similar CBD tag removed using intervening thrombin cleavage site was produced in BL21(DE3) bacteria a using pET11b vector (Novagen).

    Construct:

    Article Title: The T300A Crohn's disease risk polymorphism impairs function of the WD40 domain of ATG16L1
    Article Snippet: Loaded beads were washed extensively and used for ATG16L1 pull-down from induced crude bacterial lysates containing ATG16L1-T300 or -A300 proteins expressed from the pET plasmid (Novagen) in BL21(DE3) bacteria. .. Expression of these constructs in the soluble fraction was poor, but detectable.

    Incubation:

    Article Title: The T300A Crohn's disease risk polymorphism impairs function of the WD40 domain of ATG16L1
    Article Snippet: Lysates were cleared by centrifugation and incubated (1 h, 4 °C, rotation) with agarose beads coupled to glutathione (GE Healthcare). .. Loaded beads were washed extensively and used for ATG16L1 pull-down from induced crude bacterial lysates containing ATG16L1-T300 or -A300 proteins expressed from the pET plasmid (Novagen) in BL21(DE3) bacteria.

    Article Title: T-LAK Cell-originated Protein Kinase (TOPK) Phosphorylation of Prx1 at Ser-32 Prevents UVB-induced Apoptosis in RPMI7951 Melanoma Cells through the Regulation of Prx1 Peroxidase Activity *
    Article Snippet: His-Prx1-Wt (Wt), His-Prx1-S32A (S32A), His Prx1-S126A (S126A), His-Prx3 fusion proteins or His-Prx1-deletion mutant proteins (Ndel , Cdel or Δdel ) were expressed in BL21 (DE3) bacteria (Novagen). .. After sonication and centrifugation, the supernatant fraction was incubated with Ni-NTA-agarose beads (Qiagen, Valencia, CA) overnight at 4 °C.

    Article Title: The T-LAK Cell-originated Protein Kinase Signal Pathway Promotes Colorectal Cancer Metastasis
    Article Snippet: The His-TOPKT9A (pET46-TOPKT9A) fusion protein was expressed in BL21 (DE3) bacteria (Novagen, USA). .. After sonication and centrifugation, the supernatant fraction was incubated with Ni-NTA-agarose beads (Qiagen, Germany) overnight at 4 °C.

    Article Title: HTLV-1 Tax plugs and freezes UPF1 helicase leading to nonsense-mediated mRNA decay inhibition
    Article Snippet: .. Protein expression and purification GST-Tax proteins were produced from BL21-DE3 bacteria (Novagen) induced with 0.1 mM IPTG at 25 °C during 1 h. After sonication in MTPBS (150 mM NaCl, 12.5 mM Na2 HPO4 , 2.5 mM KH2 PO4 , 100 mM EDTA pH 7.3, 0.05% Triton,10% glycerol), the lysate was incubated for 3 h at 4 °C with glutathione magnetic beads (Promega). ..

    Article Title: HTLV-1 Tax plugs and freezes UPF1 helicase leading to nonsense-mediated mRNA decay inhibition
    Article Snippet: .. GST-Tax proteins were produced from BL21-DE3 bacteria (Novagen) induced with 0.1 mM IPTG at 25 °C during 1 h. After sonication in MTPBS (150 mM NaCl, 12.5 mM Na2 HPO4 , 2.5 mM KH2 PO4 , 100 mM EDTA pH 7.3, 0.05% Triton,10% glycerol), the lysate was incubated for 3 h at 4 °C with glutathione magnetic beads (Promega). ..

    Expressing:

    Article Title: The T-LAK Cell-originated Protein Kinase Signal Pathway Promotes Colorectal Cancer Metastasis
    Article Snippet: .. 2.5 Bacterial Expression and Purification of Fusion Proteins His-tagged proteins, including pQE81L-PRPK, pET46-WtPRPK, pET46-MutPRPK (S250A), were expressed in BL21 (DE3) bacteria (Novagen, USA) and purified as described for purification of peroxiredoxin 1 (Prx1) ( ). .. The His-TOPKT9A (pET46-TOPKT9A) fusion protein was expressed in BL21 (DE3) bacteria (Novagen, USA).

    Article Title: The T300A Crohn's disease risk polymorphism impairs function of the WD40 domain of ATG16L1
    Article Snippet: Pull-down assays Isopropyl-β-D -thiogalactoside-induced bacteria (BL21) expressing the relevant GST fusion proteins (pGEX plasmid, GE Healthcare) were lysed by treatment with lysozyme (100 μg ml−1 ), freeze-thawing and sonication in a buffer containing 20 mM Tris HCl pH 8.0, 500 mM NaCl, 1 mM EDTA and 0.1% Triton X-100 (NET buffer). .. Loaded beads were washed extensively and used for ATG16L1 pull-down from induced crude bacterial lysates containing ATG16L1-T300 or -A300 proteins expressed from the pET plasmid (Novagen) in BL21(DE3) bacteria.

    Article Title: Characterization of ubiquitin and ubiquitin-like-protein isopeptidase activities
    Article Snippet: Paragraph title: Expression and purification of UBL-PLA2 ... The mature mouse group X PLA2 (amino acids 29–151) was cloned and expressed in BL21 (DE3) bacteria (Novagen) using standard molecular biology techniques (see Supplemental Table 2 for primer information).

    Article Title: EarlyCDT(R)-Lung test: improved clinical utility through additional autoantibody assays
    Article Snippet: Antigen production Recombinant proteins were cloned into pET expression vectors (Invitrogen) and transformed into Escherichia coli BL21 (DE3) bacteria. .. The recombinant proteins were expressed in BL21 (DE3) bacteria (Novagen) and grown in terrific broth (TB), autoinduction TB media (Novagen), ECPM media, or Power Broth (Molecular Dimensions).

    Article Title: T-LAK Cell-originated Protein Kinase (TOPK) Phosphorylation of Prx1 at Ser-32 Prevents UVB-induced Apoptosis in RPMI7951 Melanoma Cells through the Regulation of Prx1 Peroxidase Activity *
    Article Snippet: Paragraph title: Bacterial Expression and Purification of His-Prx1 or Mutant Prx1 or Prx3 or Prx1 Deletion Fusion Proteins ... His-Prx1-Wt (Wt), His-Prx1-S32A (S32A), His Prx1-S126A (S126A), His-Prx3 fusion proteins or His-Prx1-deletion mutant proteins (Ndel , Cdel or Δdel ) were expressed in BL21 (DE3) bacteria (Novagen).

    Article Title: Nutrient Isothiocyanates Covalently Modify and Inhibit the Inflammatory Cytokine Macrophage Migration Inhibitory Factor (MIF)
    Article Snippet: Expression plasmids for MIF were prepared in pcDNA3 (Stratagene) by PCR amplifying the MIF coding sequence. .. Recombinant MIF with a similar CBD tag removed using intervening thrombin cleavage site was produced in BL21(DE3) bacteria a using pET11b vector (Novagen).

    Article Title: The T-LAK Cell-originated Protein Kinase Signal Pathway Promotes Colorectal Cancer Metastasis
    Article Snippet: Paragraph title: Bacterial Expression and Purification of Fusion Proteins ... The His-TOPKT9A (pET46-TOPKT9A) fusion protein was expressed in BL21 (DE3) bacteria (Novagen, USA).

    Article Title: HTLV-1 Tax plugs and freezes UPF1 helicase leading to nonsense-mediated mRNA decay inhibition
    Article Snippet: .. Protein expression and purification GST-Tax proteins were produced from BL21-DE3 bacteria (Novagen) induced with 0.1 mM IPTG at 25 °C during 1 h. After sonication in MTPBS (150 mM NaCl, 12.5 mM Na2 HPO4 , 2.5 mM KH2 PO4 , 100 mM EDTA pH 7.3, 0.05% Triton,10% glycerol), the lysate was incubated for 3 h at 4 °C with glutathione magnetic beads (Promega). ..

    Article Title: HTLV-1 Tax plugs and freezes UPF1 helicase leading to nonsense-mediated mRNA decay inhibition
    Article Snippet: Paragraph title: Protein expression and purification ... GST-Tax proteins were produced from BL21-DE3 bacteria (Novagen) induced with 0.1 mM IPTG at 25 °C during 1 h. After sonication in MTPBS (150 mM NaCl, 12.5 mM Na2 HPO4 , 2.5 mM KH2 PO4 , 100 mM EDTA pH 7.3, 0.05% Triton,10% glycerol), the lysate was incubated for 3 h at 4 °C with glutathione magnetic beads (Promega).

    Western Blot:

    Article Title: The Gos28 SNARE Protein Mediates Intra-Golgi Transport of Rhodopsin and Is Required for Photoreceptor Survival *
    Article Snippet: Paragraph title: Western Blot Analysis ... Finally, polyclonal antibodies directed to Gos28 were generated in rabbits by Cocalico Biologicals (Reamstown, PA) using a full-length Gos28 fusion protein, produced with the pET46 Ek/LIC vector and expressed in BL21(DE3) bacteria with the Novagen Overnight ExpressTM Autoinduction System 1 (EMD Biosciences, San Diego).

    Article Title: The T300A Crohn's disease risk polymorphism impairs function of the WD40 domain of ATG16L1
    Article Snippet: Loaded beads were washed extensively and used for ATG16L1 pull-down from induced crude bacterial lysates containing ATG16L1-T300 or -A300 proteins expressed from the pET plasmid (Novagen) in BL21(DE3) bacteria. .. Beads were incubated with the amount of ATG16L1-containing lysate equivalent to 1 ml of induced culture (3 h, 4 °C, rotation), washed and processed for western blotting.

    Article Title: EarlyCDT(R)-Lung test: improved clinical utility through additional autoantibody assays
    Article Snippet: The recombinant proteins were expressed in BL21 (DE3) bacteria (Novagen) and grown in terrific broth (TB), autoinduction TB media (Novagen), ECPM media, or Power Broth (Molecular Dimensions). .. Quality control tests for acceptance of protein included SDS–PAGE, Western blotting with appropriate antibodies, and analytical size exclusion chromatography.

    Transformation Assay:

    Article Title: EarlyCDT(R)-Lung test: improved clinical utility through additional autoantibody assays
    Article Snippet: Antigen production Recombinant proteins were cloned into pET expression vectors (Invitrogen) and transformed into Escherichia coli BL21 (DE3) bacteria. .. The recombinant proteins were expressed in BL21 (DE3) bacteria (Novagen) and grown in terrific broth (TB), autoinduction TB media (Novagen), ECPM media, or Power Broth (Molecular Dimensions).

    Article Title: HTLV-1 Tax plugs and freezes UPF1 helicase leading to nonsense-mediated mRNA decay inhibition
    Article Snippet: Protein expression and purification GST-Tax proteins were produced from BL21-DE3 bacteria (Novagen) induced with 0.1 mM IPTG at 25 °C during 1 h. After sonication in MTPBS (150 mM NaCl, 12.5 mM Na2 HPO4 , 2.5 mM KH2 PO4 , 100 mM EDTA pH 7.3, 0.05% Triton,10% glycerol), the lysate was incubated for 3 h at 4 °C with glutathione magnetic beads (Promega). .. Briefly, BL21 DE3 bacteria were transformed and induced overnight at 16 °C in 1 l of LB.

    Article Title: HTLV-1 Tax plugs and freezes UPF1 helicase leading to nonsense-mediated mRNA decay inhibition
    Article Snippet: GST-Tax proteins were produced from BL21-DE3 bacteria (Novagen) induced with 0.1 mM IPTG at 25 °C during 1 h. After sonication in MTPBS (150 mM NaCl, 12.5 mM Na2 HPO4 , 2.5 mM KH2 PO4 , 100 mM EDTA pH 7.3, 0.05% Triton,10% glycerol), the lysate was incubated for 3 h at 4 °C with glutathione magnetic beads (Promega). .. Briefly, BL21 DE3 bacteria were transformed and induced overnight at 16 °C in 1 l of LB.

    Cell Culture:

    Article Title: Nutrient Isothiocyanates Covalently Modify and Inhibit the Inflammatory Cytokine Macrophage Migration Inhibitory Factor (MIF)
    Article Snippet: Paragraph title: Cell Culture, Plasmids and Reagents ... Recombinant MIF with a similar CBD tag removed using intervening thrombin cleavage site was produced in BL21(DE3) bacteria a using pET11b vector (Novagen).

    Generated:

    Article Title: The Gos28 SNARE Protein Mediates Intra-Golgi Transport of Rhodopsin and Is Required for Photoreceptor Survival *
    Article Snippet: .. Finally, polyclonal antibodies directed to Gos28 were generated in rabbits by Cocalico Biologicals (Reamstown, PA) using a full-length Gos28 fusion protein, produced with the pET46 Ek/LIC vector and expressed in BL21(DE3) bacteria with the Novagen Overnight ExpressTM Autoinduction System 1 (EMD Biosciences, San Diego). .. The immunoreactive proteins were visualized using horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG (Invitrogen), goat anti-rabbit IgG (Invitrogen), donkey anti-goat IgG (Santa Cruz Biotechnology), or goat anti-mouse IgM (Jackson ImmunoResearch) followed by ECL detection (Thermo Scientific).

    Article Title: Characterization of ubiquitin and ubiquitin-like-protein isopeptidase activities
    Article Snippet: The mature mouse group X PLA2 (amino acids 29–151) was cloned and expressed in BL21 (DE3) bacteria (Novagen) using standard molecular biology techniques (see Supplemental Table 2 for primer information). .. Free catalytically active PLA2 was generated by incubating SUMO3-PLA2 with SENP2 overnight at room temperature to ensure complete cleavage prior to affinity subtraction with Ni-NTA beads.

    Polymerase Chain Reaction:

    Article Title: Nutrient Isothiocyanates Covalently Modify and Inhibit the Inflammatory Cytokine Macrophage Migration Inhibitory Factor (MIF)
    Article Snippet: Expression plasmids for MIF were prepared in pcDNA3 (Stratagene) by PCR amplifying the MIF coding sequence. .. Recombinant MIF with a similar CBD tag removed using intervening thrombin cleavage site was produced in BL21(DE3) bacteria a using pET11b vector (Novagen).

    Sonication:

    Article Title: The T300A Crohn's disease risk polymorphism impairs function of the WD40 domain of ATG16L1
    Article Snippet: Pull-down assays Isopropyl-β-D -thiogalactoside-induced bacteria (BL21) expressing the relevant GST fusion proteins (pGEX plasmid, GE Healthcare) were lysed by treatment with lysozyme (100 μg ml−1 ), freeze-thawing and sonication in a buffer containing 20 mM Tris HCl pH 8.0, 500 mM NaCl, 1 mM EDTA and 0.1% Triton X-100 (NET buffer). .. Loaded beads were washed extensively and used for ATG16L1 pull-down from induced crude bacterial lysates containing ATG16L1-T300 or -A300 proteins expressed from the pET plasmid (Novagen) in BL21(DE3) bacteria.

    Article Title: T-LAK Cell-originated Protein Kinase (TOPK) Phosphorylation of Prx1 at Ser-32 Prevents UVB-induced Apoptosis in RPMI7951 Melanoma Cells through the Regulation of Prx1 Peroxidase Activity *
    Article Snippet: His-Prx1-Wt (Wt), His-Prx1-S32A (S32A), His Prx1-S126A (S126A), His-Prx3 fusion proteins or His-Prx1-deletion mutant proteins (Ndel , Cdel or Δdel ) were expressed in BL21 (DE3) bacteria (Novagen). .. After sonication and centrifugation, the supernatant fraction was incubated with Ni-NTA-agarose beads (Qiagen, Valencia, CA) overnight at 4 °C.

    Article Title: The T-LAK Cell-originated Protein Kinase Signal Pathway Promotes Colorectal Cancer Metastasis
    Article Snippet: The His-TOPKT9A (pET46-TOPKT9A) fusion protein was expressed in BL21 (DE3) bacteria (Novagen, USA). .. After sonication and centrifugation, the supernatant fraction was incubated with Ni-NTA-agarose beads (Qiagen, Germany) overnight at 4 °C.

    Article Title: HTLV-1 Tax plugs and freezes UPF1 helicase leading to nonsense-mediated mRNA decay inhibition
    Article Snippet: .. Protein expression and purification GST-Tax proteins were produced from BL21-DE3 bacteria (Novagen) induced with 0.1 mM IPTG at 25 °C during 1 h. After sonication in MTPBS (150 mM NaCl, 12.5 mM Na2 HPO4 , 2.5 mM KH2 PO4 , 100 mM EDTA pH 7.3, 0.05% Triton,10% glycerol), the lysate was incubated for 3 h at 4 °C with glutathione magnetic beads (Promega). ..

    Article Title: HTLV-1 Tax plugs and freezes UPF1 helicase leading to nonsense-mediated mRNA decay inhibition
    Article Snippet: .. GST-Tax proteins were produced from BL21-DE3 bacteria (Novagen) induced with 0.1 mM IPTG at 25 °C during 1 h. After sonication in MTPBS (150 mM NaCl, 12.5 mM Na2 HPO4 , 2.5 mM KH2 PO4 , 100 mM EDTA pH 7.3, 0.05% Triton,10% glycerol), the lysate was incubated for 3 h at 4 °C with glutathione magnetic beads (Promega). ..

    Recombinant:

    Article Title: NC1 domain of collagen ?3(IV) derived from the basement membrane regulates Sertoli cell blood-testis barrier dynamics
    Article Snippet: .. Production of antibody against Colα3(IV) NC1 domain Recombinant Colα3(IV) NC1 domain was produced using the pET30 Ek/LIC vector kit in BL21 (DE3) bacteria (EMD Millipore). .. Colα3(IV) NC1 domain was amplified by sense 5′-GACGACGACAAGATCACAAGAATGAGAGGCTTCATCTTCA-3′ and antisense 5′-GAGGAGAAGCCCGGTGTGTCTTTTCTTCATGCACACCTGA-3′ primers using 90-d-old rat testis cDNA as template.

    Article Title: Characterization of ubiquitin and ubiquitin-like-protein isopeptidase activities
    Article Snippet: The mature mouse group X PLA2 (amino acids 29–151) was cloned and expressed in BL21 (DE3) bacteria (Novagen) using standard molecular biology techniques (see Supplemental Table 2 for primer information). .. Subsequently, PLA2 was also subcloned into the bacterial expression vectors pET24d-6His-SUMO3, pET24d-6His-NEDD8, and pET24d-6His-ISG15 to generate recombinant 6His-SUMO3-PLA2 , 6His-NEDD8-PLA2 , and 6His-ISG15-PLA2 , respectively.

    Article Title: EarlyCDT(R)-Lung test: improved clinical utility through additional autoantibody assays
    Article Snippet: .. The recombinant proteins were expressed in BL21 (DE3) bacteria (Novagen) and grown in terrific broth (TB), autoinduction TB media (Novagen), ECPM media, or Power Broth (Molecular Dimensions). .. Recombinant proteins were purified by metal chelate affinity chromatography and refolded by dialysis [ , ].

    Article Title: Nutrient Isothiocyanates Covalently Modify and Inhibit the Inflammatory Cytokine Macrophage Migration Inhibitory Factor (MIF)
    Article Snippet: .. Recombinant MIF with a similar CBD tag removed using intervening thrombin cleavage site was produced in BL21(DE3) bacteria a using pET11b vector (Novagen). ..

    Magnetic Beads:

    Article Title: HTLV-1 Tax plugs and freezes UPF1 helicase leading to nonsense-mediated mRNA decay inhibition
    Article Snippet: .. Protein expression and purification GST-Tax proteins were produced from BL21-DE3 bacteria (Novagen) induced with 0.1 mM IPTG at 25 °C during 1 h. After sonication in MTPBS (150 mM NaCl, 12.5 mM Na2 HPO4 , 2.5 mM KH2 PO4 , 100 mM EDTA pH 7.3, 0.05% Triton,10% glycerol), the lysate was incubated for 3 h at 4 °C with glutathione magnetic beads (Promega). ..

    Article Title: HTLV-1 Tax plugs and freezes UPF1 helicase leading to nonsense-mediated mRNA decay inhibition
    Article Snippet: .. GST-Tax proteins were produced from BL21-DE3 bacteria (Novagen) induced with 0.1 mM IPTG at 25 °C during 1 h. After sonication in MTPBS (150 mM NaCl, 12.5 mM Na2 HPO4 , 2.5 mM KH2 PO4 , 100 mM EDTA pH 7.3, 0.05% Triton,10% glycerol), the lysate was incubated for 3 h at 4 °C with glutathione magnetic beads (Promega). ..

    Mutagenesis:

    Article Title: T-LAK Cell-originated Protein Kinase (TOPK) Phosphorylation of Prx1 at Ser-32 Prevents UVB-induced Apoptosis in RPMI7951 Melanoma Cells through the Regulation of Prx1 Peroxidase Activity *
    Article Snippet: .. His-Prx1-Wt (Wt), His-Prx1-S32A (S32A), His Prx1-S126A (S126A), His-Prx3 fusion proteins or His-Prx1-deletion mutant proteins (Ndel , Cdel or Δdel ) were expressed in BL21 (DE3) bacteria (Novagen). .. Bacteria were grown at 37 °C to an absorbance of 0.9–1.0 at 600 nm and induced with 0.5 m m IPTG overnight at 15 °C and harvested by centrifugation.

    Size-exclusion Chromatography:

    Article Title: EarlyCDT(R)-Lung test: improved clinical utility through additional autoantibody assays
    Article Snippet: The recombinant proteins were expressed in BL21 (DE3) bacteria (Novagen) and grown in terrific broth (TB), autoinduction TB media (Novagen), ECPM media, or Power Broth (Molecular Dimensions). .. Quality control tests for acceptance of protein included SDS–PAGE, Western blotting with appropriate antibodies, and analytical size exclusion chromatography.

    Article Title: MICU1 imparts the mitochondrial uniporter with the ability to discriminate between Ca2+ and Mn2+
    Article Snippet: Briefly, proteins were expressed in BL21* (DE3) bacteria (Millipore) and subjected to Ni-NTA purification. .. The proteins were further purified by size exclusion chromatography (SEC) using a Superdex 200 column and buffer A (25 mM Hepes, pH 7, and 150 mM NaCl) along with 0.5 mM EGTA.

    Purification:

    Article Title: The T-LAK Cell-originated Protein Kinase Signal Pathway Promotes Colorectal Cancer Metastasis
    Article Snippet: .. 2.5 Bacterial Expression and Purification of Fusion Proteins His-tagged proteins, including pQE81L-PRPK, pET46-WtPRPK, pET46-MutPRPK (S250A), were expressed in BL21 (DE3) bacteria (Novagen, USA) and purified as described for purification of peroxiredoxin 1 (Prx1) ( ). .. The His-TOPKT9A (pET46-TOPKT9A) fusion protein was expressed in BL21 (DE3) bacteria (Novagen, USA).

    Article Title: Characterization of ubiquitin and ubiquitin-like-protein isopeptidase activities
    Article Snippet: Paragraph title: Expression and purification of UBL-PLA2 ... The mature mouse group X PLA2 (amino acids 29–151) was cloned and expressed in BL21 (DE3) bacteria (Novagen) using standard molecular biology techniques (see Supplemental Table 2 for primer information).

    Article Title: EarlyCDT(R)-Lung test: improved clinical utility through additional autoantibody assays
    Article Snippet: The recombinant proteins were expressed in BL21 (DE3) bacteria (Novagen) and grown in terrific broth (TB), autoinduction TB media (Novagen), ECPM media, or Power Broth (Molecular Dimensions). .. Recombinant proteins were purified by metal chelate affinity chromatography and refolded by dialysis [ , ].

    Article Title: MICU1 imparts the mitochondrial uniporter with the ability to discriminate between Ca2+ and Mn2+
    Article Snippet: .. Briefly, proteins were expressed in BL21* (DE3) bacteria (Millipore) and subjected to Ni-NTA purification. .. The proteins were further purified by size exclusion chromatography (SEC) using a Superdex 200 column and buffer A (25 mM Hepes, pH 7, and 150 mM NaCl) along with 0.5 mM EGTA.

    Article Title: T-LAK Cell-originated Protein Kinase (TOPK) Phosphorylation of Prx1 at Ser-32 Prevents UVB-induced Apoptosis in RPMI7951 Melanoma Cells through the Regulation of Prx1 Peroxidase Activity *
    Article Snippet: Paragraph title: Bacterial Expression and Purification of His-Prx1 or Mutant Prx1 or Prx3 or Prx1 Deletion Fusion Proteins ... His-Prx1-Wt (Wt), His-Prx1-S32A (S32A), His Prx1-S126A (S126A), His-Prx3 fusion proteins or His-Prx1-deletion mutant proteins (Ndel , Cdel or Δdel ) were expressed in BL21 (DE3) bacteria (Novagen).

    Article Title: Nutrient Isothiocyanates Covalently Modify and Inhibit the Inflammatory Cytokine Macrophage Migration Inhibitory Factor (MIF)
    Article Snippet: In some cases, the MIF coding region was fused to a chitin binding domain (CBD) tag at the C terminus for purification on chitin beads. .. Recombinant MIF with a similar CBD tag removed using intervening thrombin cleavage site was produced in BL21(DE3) bacteria a using pET11b vector (Novagen).

    Article Title: The T-LAK Cell-originated Protein Kinase Signal Pathway Promotes Colorectal Cancer Metastasis
    Article Snippet: Paragraph title: Bacterial Expression and Purification of Fusion Proteins ... The His-TOPKT9A (pET46-TOPKT9A) fusion protein was expressed in BL21 (DE3) bacteria (Novagen, USA).

    Article Title: HTLV-1 Tax plugs and freezes UPF1 helicase leading to nonsense-mediated mRNA decay inhibition
    Article Snippet: .. Protein expression and purification GST-Tax proteins were produced from BL21-DE3 bacteria (Novagen) induced with 0.1 mM IPTG at 25 °C during 1 h. After sonication in MTPBS (150 mM NaCl, 12.5 mM Na2 HPO4 , 2.5 mM KH2 PO4 , 100 mM EDTA pH 7.3, 0.05% Triton,10% glycerol), the lysate was incubated for 3 h at 4 °C with glutathione magnetic beads (Promega). ..

    Article Title: HTLV-1 Tax plugs and freezes UPF1 helicase leading to nonsense-mediated mRNA decay inhibition
    Article Snippet: Paragraph title: Protein expression and purification ... GST-Tax proteins were produced from BL21-DE3 bacteria (Novagen) induced with 0.1 mM IPTG at 25 °C during 1 h. After sonication in MTPBS (150 mM NaCl, 12.5 mM Na2 HPO4 , 2.5 mM KH2 PO4 , 100 mM EDTA pH 7.3, 0.05% Triton,10% glycerol), the lysate was incubated for 3 h at 4 °C with glutathione magnetic beads (Promega).

    Protein Purification:

    Article Title: MICU1 imparts the mitochondrial uniporter with the ability to discriminate between Ca2+ and Mn2+
    Article Snippet: Paragraph title: Protein Purification. ... Briefly, proteins were expressed in BL21* (DE3) bacteria (Millipore) and subjected to Ni-NTA purification.

    Sequencing:

    Article Title: NC1 domain of collagen ?3(IV) derived from the basement membrane regulates Sertoli cell blood-testis barrier dynamics
    Article Snippet: Production of antibody against Colα3(IV) NC1 domain Recombinant Colα3(IV) NC1 domain was produced using the pET30 Ek/LIC vector kit in BL21 (DE3) bacteria (EMD Millipore). .. The stop codon in the original full-length sequence was omitted due to the presence of in frame stop codon in the vector.

    Article Title: Nutrient Isothiocyanates Covalently Modify and Inhibit the Inflammatory Cytokine Macrophage Migration Inhibitory Factor (MIF)
    Article Snippet: Expression plasmids for MIF were prepared in pcDNA3 (Stratagene) by PCR amplifying the MIF coding sequence. .. Recombinant MIF with a similar CBD tag removed using intervening thrombin cleavage site was produced in BL21(DE3) bacteria a using pET11b vector (Novagen).

    Affinity Chromatography:

    Article Title: EarlyCDT(R)-Lung test: improved clinical utility through additional autoantibody assays
    Article Snippet: The recombinant proteins were expressed in BL21 (DE3) bacteria (Novagen) and grown in terrific broth (TB), autoinduction TB media (Novagen), ECPM media, or Power Broth (Molecular Dimensions). .. Recombinant proteins were purified by metal chelate affinity chromatography and refolded by dialysis [ , ].

    SDS Page:

    Article Title: EarlyCDT(R)-Lung test: improved clinical utility through additional autoantibody assays
    Article Snippet: The recombinant proteins were expressed in BL21 (DE3) bacteria (Novagen) and grown in terrific broth (TB), autoinduction TB media (Novagen), ECPM media, or Power Broth (Molecular Dimensions). .. Quality control tests for acceptance of protein included SDS–PAGE, Western blotting with appropriate antibodies, and analytical size exclusion chromatography.

    Plasmid Preparation:

    Article Title: The Gos28 SNARE Protein Mediates Intra-Golgi Transport of Rhodopsin and Is Required for Photoreceptor Survival *
    Article Snippet: .. Finally, polyclonal antibodies directed to Gos28 were generated in rabbits by Cocalico Biologicals (Reamstown, PA) using a full-length Gos28 fusion protein, produced with the pET46 Ek/LIC vector and expressed in BL21(DE3) bacteria with the Novagen Overnight ExpressTM Autoinduction System 1 (EMD Biosciences, San Diego). .. The immunoreactive proteins were visualized using horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG (Invitrogen), goat anti-rabbit IgG (Invitrogen), donkey anti-goat IgG (Santa Cruz Biotechnology), or goat anti-mouse IgM (Jackson ImmunoResearch) followed by ECL detection (Thermo Scientific).

    Article Title: NC1 domain of collagen ?3(IV) derived from the basement membrane regulates Sertoli cell blood-testis barrier dynamics
    Article Snippet: .. Production of antibody against Colα3(IV) NC1 domain Recombinant Colα3(IV) NC1 domain was produced using the pET30 Ek/LIC vector kit in BL21 (DE3) bacteria (EMD Millipore). .. Colα3(IV) NC1 domain was amplified by sense 5′-GACGACGACAAGATCACAAGAATGAGAGGCTTCATCTTCA-3′ and antisense 5′-GAGGAGAAGCCCGGTGTGTCTTTTCTTCATGCACACCTGA-3′ primers using 90-d-old rat testis cDNA as template.

    Article Title: The T300A Crohn's disease risk polymorphism impairs function of the WD40 domain of ATG16L1
    Article Snippet: .. Loaded beads were washed extensively and used for ATG16L1 pull-down from induced crude bacterial lysates containing ATG16L1-T300 or -A300 proteins expressed from the pET plasmid (Novagen) in BL21(DE3) bacteria. .. Expression of these constructs in the soluble fraction was poor, but detectable.

    Article Title: Nutrient Isothiocyanates Covalently Modify and Inhibit the Inflammatory Cytokine Macrophage Migration Inhibitory Factor (MIF)
    Article Snippet: .. Recombinant MIF with a similar CBD tag removed using intervening thrombin cleavage site was produced in BL21(DE3) bacteria a using pET11b vector (Novagen). ..

    Negative Control:

    Article Title: EarlyCDT(R)-Lung test: improved clinical utility through additional autoantibody assays
    Article Snippet: Negative control proteins were also produced (BirA and NusA tags alone). .. The recombinant proteins were expressed in BL21 (DE3) bacteria (Novagen) and grown in terrific broth (TB), autoinduction TB media (Novagen), ECPM media, or Power Broth (Molecular Dimensions).

    Positron Emission Tomography:

    Article Title: The T300A Crohn's disease risk polymorphism impairs function of the WD40 domain of ATG16L1
    Article Snippet: .. Loaded beads were washed extensively and used for ATG16L1 pull-down from induced crude bacterial lysates containing ATG16L1-T300 or -A300 proteins expressed from the pET plasmid (Novagen) in BL21(DE3) bacteria. .. Expression of these constructs in the soluble fraction was poor, but detectable.

    Article Title: EarlyCDT(R)-Lung test: improved clinical utility through additional autoantibody assays
    Article Snippet: Antigen production Recombinant proteins were cloned into pET expression vectors (Invitrogen) and transformed into Escherichia coli BL21 (DE3) bacteria. .. The recombinant proteins were expressed in BL21 (DE3) bacteria (Novagen) and grown in terrific broth (TB), autoinduction TB media (Novagen), ECPM media, or Power Broth (Molecular Dimensions).

    Produced:

    Article Title: The Gos28 SNARE Protein Mediates Intra-Golgi Transport of Rhodopsin and Is Required for Photoreceptor Survival *
    Article Snippet: .. Finally, polyclonal antibodies directed to Gos28 were generated in rabbits by Cocalico Biologicals (Reamstown, PA) using a full-length Gos28 fusion protein, produced with the pET46 Ek/LIC vector and expressed in BL21(DE3) bacteria with the Novagen Overnight ExpressTM Autoinduction System 1 (EMD Biosciences, San Diego). .. The immunoreactive proteins were visualized using horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG (Invitrogen), goat anti-rabbit IgG (Invitrogen), donkey anti-goat IgG (Santa Cruz Biotechnology), or goat anti-mouse IgM (Jackson ImmunoResearch) followed by ECL detection (Thermo Scientific).

    Article Title: NC1 domain of collagen ?3(IV) derived from the basement membrane regulates Sertoli cell blood-testis barrier dynamics
    Article Snippet: .. Production of antibody against Colα3(IV) NC1 domain Recombinant Colα3(IV) NC1 domain was produced using the pET30 Ek/LIC vector kit in BL21 (DE3) bacteria (EMD Millipore). .. Colα3(IV) NC1 domain was amplified by sense 5′-GACGACGACAAGATCACAAGAATGAGAGGCTTCATCTTCA-3′ and antisense 5′-GAGGAGAAGCCCGGTGTGTCTTTTCTTCATGCACACCTGA-3′ primers using 90-d-old rat testis cDNA as template.

    Article Title: EarlyCDT(R)-Lung test: improved clinical utility through additional autoantibody assays
    Article Snippet: Negative control proteins were also produced (BirA and NusA tags alone). .. The recombinant proteins were expressed in BL21 (DE3) bacteria (Novagen) and grown in terrific broth (TB), autoinduction TB media (Novagen), ECPM media, or Power Broth (Molecular Dimensions).

    Article Title: Nutrient Isothiocyanates Covalently Modify and Inhibit the Inflammatory Cytokine Macrophage Migration Inhibitory Factor (MIF)
    Article Snippet: .. Recombinant MIF with a similar CBD tag removed using intervening thrombin cleavage site was produced in BL21(DE3) bacteria a using pET11b vector (Novagen). ..

    Article Title: HTLV-1 Tax plugs and freezes UPF1 helicase leading to nonsense-mediated mRNA decay inhibition
    Article Snippet: .. Protein expression and purification GST-Tax proteins were produced from BL21-DE3 bacteria (Novagen) induced with 0.1 mM IPTG at 25 °C during 1 h. After sonication in MTPBS (150 mM NaCl, 12.5 mM Na2 HPO4 , 2.5 mM KH2 PO4 , 100 mM EDTA pH 7.3, 0.05% Triton,10% glycerol), the lysate was incubated for 3 h at 4 °C with glutathione magnetic beads (Promega). ..

    Article Title: HTLV-1 Tax plugs and freezes UPF1 helicase leading to nonsense-mediated mRNA decay inhibition
    Article Snippet: .. GST-Tax proteins were produced from BL21-DE3 bacteria (Novagen) induced with 0.1 mM IPTG at 25 °C during 1 h. After sonication in MTPBS (150 mM NaCl, 12.5 mM Na2 HPO4 , 2.5 mM KH2 PO4 , 100 mM EDTA pH 7.3, 0.05% Triton,10% glycerol), the lysate was incubated for 3 h at 4 °C with glutathione magnetic beads (Promega). ..

    Lysis:

    Article Title: The T-LAK Cell-originated Protein Kinase Signal Pathway Promotes Colorectal Cancer Metastasis
    Article Snippet: 2.5 Bacterial Expression and Purification of Fusion Proteins His-tagged proteins, including pQE81L-PRPK, pET46-WtPRPK, pET46-MutPRPK (S250A), were expressed in BL21 (DE3) bacteria (Novagen, USA) and purified as described for purification of peroxiredoxin 1 (Prx1) ( ). .. Cell pellets were suspended in 50 mM Tris (pH 8.0) lysis buffer containing 200 mM NaCl and 10 mM imidazole.

    Article Title: T-LAK Cell-originated Protein Kinase (TOPK) Phosphorylation of Prx1 at Ser-32 Prevents UVB-induced Apoptosis in RPMI7951 Melanoma Cells through the Regulation of Prx1 Peroxidase Activity *
    Article Snippet: His-Prx1-Wt (Wt), His-Prx1-S32A (S32A), His Prx1-S126A (S126A), His-Prx3 fusion proteins or His-Prx1-deletion mutant proteins (Ndel , Cdel or Δdel ) were expressed in BL21 (DE3) bacteria (Novagen). .. Cell pellets were suspended in 50 m m Tris-HCl (pH 8.0) lysis buffer containing 200 m m NaCl, 10 m m imidazole, and 10 m m mercaptoethanol.

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    Millipore strain bl21
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