novablue de3 competent e  (Millipore)


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    Structured Review

    Millipore novablue de3 competent e
    Novablue De3 Competent E, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/novablue de3 competent e/product/Millipore
    Average 92 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    novablue de3 competent e - by Bioz Stars, 2020-04
    92/100 stars

    Related Products / Commonly Used Together

    sfβgly
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    Related Articles

    Clone Assay:

    Article Title: Using the Amino Acid Network to Modulate the Hydrolytic Activity of β-Glycosidases
    Article Snippet: .. Wild-type or mutants of Sfβgly cloned into pAE were used to transform NovaBlue (DE3) competent E . coli cells (EMD Millipore, Billerica, MA, USA). ..

    Article Title: Orientia tsutsugamushi Strain Ikeda Ankyrin Repeat-Containing Proteins Recruit SCF1 Ubiquitin Ligase Machinery via Poxvirus-Like F-Box Motifs
    Article Snippet: Recombinant O. tsutsugamushi outer membrane protein A (OmpA; OTT_1320) was generated using primers 5′- GACGACGACAAGATA TGTTTATGGCAAAGATCTAAACATAGTAAC-3′ and 5′- GAGGAGAAGCCCGGTTA TTTATGTTTCCCATGTATAGCTTGTAAAAACTG-3′ (sequences that are compatible with ligation-independent cloning are italicized) to amplify the region corresponding to amino acids 22 to 93, which is unique to OmpA, as assessed by NCBI BLAST (Basic Local Alignment Search Tool) ( ) searches. .. Plasmids were propagated in Escherichia coli DE3 NovaBlue cells (EMD Millipore).

    Centrifugation:

    Article Title: Sets of Covariant Residues Modulate the Activity and Thermal Stability of GH1 ?-Glucosidases
    Article Snippet: Expression and purification of recombinant and wild-type Sfβgly NovaBlue (DE3) competent cells (EMD Millipore, Billerica, MA, USA) were transformed with pAE plasmids encoding wild-type or mutant Sfβgly, plated on LB-agar containing ampicillin (50 µg/mL) and grown at 37°C for 16 h. Single colonies were grown at 20°C in LB broth containing ampicillin (50 µg/mL) until they reached an attenuance of 0.500 at 600 nm. .. Next, 0.4 mM (final concentration) isopropyl β-D-1-thiogalactopyranoside (IPTG) was added for 16 h to induce recombinant protein expression, after which cells were harvested by centrifugation (4,000×g , 20 min, 4°C) and frozen at −80°C.

    Article Title: Using the Amino Acid Network to Modulate the Hydrolytic Activity of β-Glycosidases
    Article Snippet: Wild-type or mutants of Sfβgly cloned into pAE were used to transform NovaBlue (DE3) competent E . coli cells (EMD Millipore, Billerica, MA, USA). .. Cells expressing recombinant enzymes were collected by centrifugation at 4,000 × g for 20 min (4°C) and frozen at -80°C until use.

    Amplification:

    Article Title: Orientia tsutsugamushi Strain Ikeda Ankyrin Repeat-Containing Proteins Recruit SCF1 Ubiquitin Ligase Machinery via Poxvirus-Like F-Box Motifs
    Article Snippet: The primers harbored ligase-independent cloning (LIC) tails, and the amplicon was annealed with pET46 Ek/LIC (Novagen, EMD Millipore, Billerica, MA). .. Plasmids were propagated in Escherichia coli DE3 NovaBlue cells (EMD Millipore).

    Ligation:

    Article Title: Orientia tsutsugamushi Strain Ikeda Ankyrin Repeat-Containing Proteins Recruit SCF1 Ubiquitin Ligase Machinery via Poxvirus-Like F-Box Motifs
    Article Snippet: Recombinant O. tsutsugamushi outer membrane protein A (OmpA; OTT_1320) was generated using primers 5′- GACGACGACAAGATA TGTTTATGGCAAAGATCTAAACATAGTAAC-3′ and 5′- GAGGAGAAGCCCGGTTA TTTATGTTTCCCATGTATAGCTTGTAAAAACTG-3′ (sequences that are compatible with ligation-independent cloning are italicized) to amplify the region corresponding to amino acids 22 to 93, which is unique to OmpA, as assessed by NCBI BLAST (Basic Local Alignment Search Tool) ( ) searches. .. Plasmids were propagated in Escherichia coli DE3 NovaBlue cells (EMD Millipore).

    Mutagenesis:

    Article Title: Sets of Covariant Residues Modulate the Activity and Thermal Stability of GH1 ?-Glucosidases
    Article Snippet: .. Expression and purification of recombinant and wild-type Sfβgly NovaBlue (DE3) competent cells (EMD Millipore, Billerica, MA, USA) were transformed with pAE plasmids encoding wild-type or mutant Sfβgly, plated on LB-agar containing ampicillin (50 µg/mL) and grown at 37°C for 16 h. Single colonies were grown at 20°C in LB broth containing ampicillin (50 µg/mL) until they reached an attenuance of 0.500 at 600 nm. .. Next, 0.4 mM (final concentration) isopropyl β-D-1-thiogalactopyranoside (IPTG) was added for 16 h to induce recombinant protein expression, after which cells were harvested by centrifugation (4,000×g , 20 min, 4°C) and frozen at −80°C.

    Article Title: Using the Amino Acid Network to Modulate the Hydrolytic Activity of β-Glycosidases
    Article Snippet: Paragraph title: Mutagenesis, expression in E . coli and purification of wild-type Sfβgly and its mutants ... Wild-type or mutants of Sfβgly cloned into pAE were used to transform NovaBlue (DE3) competent E . coli cells (EMD Millipore, Billerica, MA, USA).

    Ligase Independent Cloning:

    Article Title: Orientia tsutsugamushi Strain Ikeda Ankyrin Repeat-Containing Proteins Recruit SCF1 Ubiquitin Ligase Machinery via Poxvirus-Like F-Box Motifs
    Article Snippet: The primers harbored ligase-independent cloning (LIC) tails, and the amplicon was annealed with pET46 Ek/LIC (Novagen, EMD Millipore, Billerica, MA). .. Plasmids were propagated in Escherichia coli DE3 NovaBlue cells (EMD Millipore).

    Incubation:

    Article Title: Sets of Covariant Residues Modulate the Activity and Thermal Stability of GH1 ?-Glucosidases
    Article Snippet: Expression and purification of recombinant and wild-type Sfβgly NovaBlue (DE3) competent cells (EMD Millipore, Billerica, MA, USA) were transformed with pAE plasmids encoding wild-type or mutant Sfβgly, plated on LB-agar containing ampicillin (50 µg/mL) and grown at 37°C for 16 h. Single colonies were grown at 20°C in LB broth containing ampicillin (50 µg/mL) until they reached an attenuance of 0.500 at 600 nm. .. The supernatants were recovered by centrifugation (13,200 g, 20 min, 4°C), and soluble recombinant proteins were incubated with 200 µL Ni-NTA Agarose (4°C, 1 h) (Qiagen, Hilden, Germany).

    Purification:

    Article Title: Sets of Covariant Residues Modulate the Activity and Thermal Stability of GH1 ?-Glucosidases
    Article Snippet: .. Expression and purification of recombinant and wild-type Sfβgly NovaBlue (DE3) competent cells (EMD Millipore, Billerica, MA, USA) were transformed with pAE plasmids encoding wild-type or mutant Sfβgly, plated on LB-agar containing ampicillin (50 µg/mL) and grown at 37°C for 16 h. Single colonies were grown at 20°C in LB broth containing ampicillin (50 µg/mL) until they reached an attenuance of 0.500 at 600 nm. .. Next, 0.4 mM (final concentration) isopropyl β-D-1-thiogalactopyranoside (IPTG) was added for 16 h to induce recombinant protein expression, after which cells were harvested by centrifugation (4,000×g , 20 min, 4°C) and frozen at −80°C.

    Article Title: Using the Amino Acid Network to Modulate the Hydrolytic Activity of β-Glycosidases
    Article Snippet: Paragraph title: Mutagenesis, expression in E . coli and purification of wild-type Sfβgly and its mutants ... Wild-type or mutants of Sfβgly cloned into pAE were used to transform NovaBlue (DE3) competent E . coli cells (EMD Millipore, Billerica, MA, USA).

    Article Title: Orientia tsutsugamushi Strain Ikeda Ankyrin Repeat-Containing Proteins Recruit SCF1 Ubiquitin Ligase Machinery via Poxvirus-Like F-Box Motifs
    Article Snippet: Plasmids were propagated in Escherichia coli DE3 NovaBlue cells (EMD Millipore). .. The final recombinant protein, which carried an N-terminal 6×His tag of 1.7 kDa, was purified ( > 95% homogeneity) by immobilized metal affinity chromatography (IMAC) (HisTrap; GE Healthcare Biosciences, Pittsburgh, PA) according to the manufacturer's protocol and concentrated using Amicon Ultra filters (EMD Millipore).

    Acid Assay:

    Article Title: Orientia tsutsugamushi Strain Ikeda Ankyrin Repeat-Containing Proteins Recruit SCF1 Ubiquitin Ligase Machinery via Poxvirus-Like F-Box Motifs
    Article Snippet: Plasmids were propagated in Escherichia coli DE3 NovaBlue cells (EMD Millipore). .. The protein concentration was determined using the bicinchoninic acid assay.

    Concentration Assay:

    Article Title: Sets of Covariant Residues Modulate the Activity and Thermal Stability of GH1 ?-Glucosidases
    Article Snippet: Expression and purification of recombinant and wild-type Sfβgly NovaBlue (DE3) competent cells (EMD Millipore, Billerica, MA, USA) were transformed with pAE plasmids encoding wild-type or mutant Sfβgly, plated on LB-agar containing ampicillin (50 µg/mL) and grown at 37°C for 16 h. Single colonies were grown at 20°C in LB broth containing ampicillin (50 µg/mL) until they reached an attenuance of 0.500 at 600 nm. .. Next, 0.4 mM (final concentration) isopropyl β-D-1-thiogalactopyranoside (IPTG) was added for 16 h to induce recombinant protein expression, after which cells were harvested by centrifugation (4,000×g , 20 min, 4°C) and frozen at −80°C.

    Generated:

    Article Title: Orientia tsutsugamushi Strain Ikeda Ankyrin Repeat-Containing Proteins Recruit SCF1 Ubiquitin Ligase Machinery via Poxvirus-Like F-Box Motifs
    Article Snippet: Recombinant O. tsutsugamushi outer membrane protein A (OmpA; OTT_1320) was generated using primers 5′- GACGACGACAAGATA TGTTTATGGCAAAGATCTAAACATAGTAAC-3′ and 5′- GAGGAGAAGCCCGGTTA TTTATGTTTCCCATGTATAGCTTGTAAAAACTG-3′ (sequences that are compatible with ligation-independent cloning are italicized) to amplify the region corresponding to amino acids 22 to 93, which is unique to OmpA, as assessed by NCBI BLAST (Basic Local Alignment Search Tool) ( ) searches. .. Plasmids were propagated in Escherichia coli DE3 NovaBlue cells (EMD Millipore).

    Affinity Chromatography:

    Article Title: Orientia tsutsugamushi Strain Ikeda Ankyrin Repeat-Containing Proteins Recruit SCF1 Ubiquitin Ligase Machinery via Poxvirus-Like F-Box Motifs
    Article Snippet: Plasmids were propagated in Escherichia coli DE3 NovaBlue cells (EMD Millipore). .. The final recombinant protein, which carried an N-terminal 6×His tag of 1.7 kDa, was purified ( > 95% homogeneity) by immobilized metal affinity chromatography (IMAC) (HisTrap; GE Healthcare Biosciences, Pittsburgh, PA) according to the manufacturer's protocol and concentrated using Amicon Ultra filters (EMD Millipore).

    Expressing:

    Article Title: Sets of Covariant Residues Modulate the Activity and Thermal Stability of GH1 ?-Glucosidases
    Article Snippet: .. Expression and purification of recombinant and wild-type Sfβgly NovaBlue (DE3) competent cells (EMD Millipore, Billerica, MA, USA) were transformed with pAE plasmids encoding wild-type or mutant Sfβgly, plated on LB-agar containing ampicillin (50 µg/mL) and grown at 37°C for 16 h. Single colonies were grown at 20°C in LB broth containing ampicillin (50 µg/mL) until they reached an attenuance of 0.500 at 600 nm. .. Next, 0.4 mM (final concentration) isopropyl β-D-1-thiogalactopyranoside (IPTG) was added for 16 h to induce recombinant protein expression, after which cells were harvested by centrifugation (4,000×g , 20 min, 4°C) and frozen at −80°C.

    Article Title: Using the Amino Acid Network to Modulate the Hydrolytic Activity of β-Glycosidases
    Article Snippet: Paragraph title: Mutagenesis, expression in E . coli and purification of wild-type Sfβgly and its mutants ... Wild-type or mutants of Sfβgly cloned into pAE were used to transform NovaBlue (DE3) competent E . coli cells (EMD Millipore, Billerica, MA, USA).

    Article Title: Orientia tsutsugamushi Strain Ikeda Ankyrin Repeat-Containing Proteins Recruit SCF1 Ubiquitin Ligase Machinery via Poxvirus-Like F-Box Motifs
    Article Snippet: Plasmids were propagated in Escherichia coli DE3 NovaBlue cells (EMD Millipore). .. Protein was expressed by inoculation of 100 ml Luria-Burtani (LB) broth supplemented with 100 mg liter−1 of ampicillin with a colony from freshly plated E. coli , and the culture was allowed to autoinduce protein expression overnight.

    Protein Concentration:

    Article Title: Orientia tsutsugamushi Strain Ikeda Ankyrin Repeat-Containing Proteins Recruit SCF1 Ubiquitin Ligase Machinery via Poxvirus-Like F-Box Motifs
    Article Snippet: Plasmids were propagated in Escherichia coli DE3 NovaBlue cells (EMD Millipore). .. The protein concentration was determined using the bicinchoninic acid assay.

    Polymerase Chain Reaction:

    Article Title: Using the Amino Acid Network to Modulate the Hydrolytic Activity of β-Glycosidases
    Article Snippet: Mutagenesis, expression in E . coli and purification of wild-type Sfβgly and its mutants The wild-type Sfβgly gene cloned into pAE plasmid [ ] was used as template for mutagenic PCR reactions using mutagenic primers (Table A in ) and the QuikChange site-directed mutagenesis kit (Stratagene, La Jolla, CA, USA), following the manufacturer’s instructions. .. Wild-type or mutants of Sfβgly cloned into pAE were used to transform NovaBlue (DE3) competent E . coli cells (EMD Millipore, Billerica, MA, USA).

    Transformation Assay:

    Article Title: Sets of Covariant Residues Modulate the Activity and Thermal Stability of GH1 ?-Glucosidases
    Article Snippet: .. Expression and purification of recombinant and wild-type Sfβgly NovaBlue (DE3) competent cells (EMD Millipore, Billerica, MA, USA) were transformed with pAE plasmids encoding wild-type or mutant Sfβgly, plated on LB-agar containing ampicillin (50 µg/mL) and grown at 37°C for 16 h. Single colonies were grown at 20°C in LB broth containing ampicillin (50 µg/mL) until they reached an attenuance of 0.500 at 600 nm. .. Next, 0.4 mM (final concentration) isopropyl β-D-1-thiogalactopyranoside (IPTG) was added for 16 h to induce recombinant protein expression, after which cells were harvested by centrifugation (4,000×g , 20 min, 4°C) and frozen at −80°C.

    Recombinant:

    Article Title: Sets of Covariant Residues Modulate the Activity and Thermal Stability of GH1 ?-Glucosidases
    Article Snippet: .. Expression and purification of recombinant and wild-type Sfβgly NovaBlue (DE3) competent cells (EMD Millipore, Billerica, MA, USA) were transformed with pAE plasmids encoding wild-type or mutant Sfβgly, plated on LB-agar containing ampicillin (50 µg/mL) and grown at 37°C for 16 h. Single colonies were grown at 20°C in LB broth containing ampicillin (50 µg/mL) until they reached an attenuance of 0.500 at 600 nm. .. Next, 0.4 mM (final concentration) isopropyl β-D-1-thiogalactopyranoside (IPTG) was added for 16 h to induce recombinant protein expression, after which cells were harvested by centrifugation (4,000×g , 20 min, 4°C) and frozen at −80°C.

    Article Title: Using the Amino Acid Network to Modulate the Hydrolytic Activity of β-Glycosidases
    Article Snippet: Wild-type or mutants of Sfβgly cloned into pAE were used to transform NovaBlue (DE3) competent E . coli cells (EMD Millipore, Billerica, MA, USA). .. Individual colonies were used to inoculate LB broth containing 50 μg/mL ampicillin and grown to an optical density of 0.5 at 600 nm, when 0.4 mM isopropyl β-D-1-thiogalactopyranoside was added to induce the expression of recombinant wild-type and mutants of Sfβgly for 16 h at 20°C.

    Article Title: Orientia tsutsugamushi Strain Ikeda Ankyrin Repeat-Containing Proteins Recruit SCF1 Ubiquitin Ligase Machinery via Poxvirus-Like F-Box Motifs
    Article Snippet: Recombinant O. tsutsugamushi outer membrane protein A (OmpA; OTT_1320) was generated using primers 5′- GACGACGACAAGATA TGTTTATGGCAAAGATCTAAACATAGTAAC-3′ and 5′- GAGGAGAAGCCCGGTTA TTTATGTTTCCCATGTATAGCTTGTAAAAACTG-3′ (sequences that are compatible with ligation-independent cloning are italicized) to amplify the region corresponding to amino acids 22 to 93, which is unique to OmpA, as assessed by NCBI BLAST (Basic Local Alignment Search Tool) ( ) searches. .. Plasmids were propagated in Escherichia coli DE3 NovaBlue cells (EMD Millipore).

    Affinity Purification:

    Article Title: Orientia tsutsugamushi Strain Ikeda Ankyrin Repeat-Containing Proteins Recruit SCF1 Ubiquitin Ligase Machinery via Poxvirus-Like F-Box Motifs
    Article Snippet: Affinity-purified rabbit polyclonal antiserum targeting Ank9 amino acids 12 to 28 was generated by New England Peptide (Gardner, MA). .. Plasmids were propagated in Escherichia coli DE3 NovaBlue cells (EMD Millipore).

    Plasmid Preparation:

    Article Title: Using the Amino Acid Network to Modulate the Hydrolytic Activity of β-Glycosidases
    Article Snippet: Mutagenesis, expression in E . coli and purification of wild-type Sfβgly and its mutants The wild-type Sfβgly gene cloned into pAE plasmid [ ] was used as template for mutagenic PCR reactions using mutagenic primers (Table A in ) and the QuikChange site-directed mutagenesis kit (Stratagene, La Jolla, CA, USA), following the manufacturer’s instructions. .. Wild-type or mutants of Sfβgly cloned into pAE were used to transform NovaBlue (DE3) competent E . coli cells (EMD Millipore, Billerica, MA, USA).

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  • 92
    Millipore novablue de3 competent e
    Novablue De3 Competent E, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/novablue de3 competent e/product/Millipore
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    Millipore wild type sfβgly novablue de3 competent cells
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