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imidazole  (Valiant Co Ltd)

 
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    Structured Review

    Valiant Co Ltd imidazole
    Imidazole, supplied by Valiant Co Ltd, used in various techniques. Bioz Stars score: 94/100, based on 62 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/02102033%2E1/Imidazole/10__1107_slash_s0907444913015771-46-11-18
    Average 94 stars, based on 62 article reviews
    imidazole - by Bioz Stars, 2026-09
    94/100 stars

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    Related Articles

    Construct:

    Article Title: Supplementary Information Unraveling the Mechanics of a Repeat-Protein Nanospring — From Folding of Individual Repeats to Fluctuations of the Superhelix
    Article Snippet: The protein was then further purified by size exclusion chromatography using a HiLoad 26/600 Superdex 75 pg or HiLoad 16/600 Superdex 75 pg (GE Healthcare) equilibrated in either Tris or phosphate buffer (50 mM Tris-HCl pH 7.5 or 50 mM sodium phosphate pH 6.8, 150 mM NaCl). .. After elution from the resin with buffer containing imidazole, the protein was dialysed against 50 mM sodium phosphate pH 6.8, 150 mM NaCl for 18 hours, in the presence of thrombin (MP biomedical) to remove the H6-tag from the construct. .. The protein was further purified using a HiLoad 26/600 Superdex 75 pg column (GE Healthcare) equilibrated in 10 (10 mM HEPES pH 7.5, 150 mM NaCl, and concentrated to 20 mg/mL.

    Expressing:

    Article Title: Structural Basis of Redox-Dependent Affinities of Dihydroorotate Dehydrogenase for Its Substrates and Products.
    Article Snippet: An Escherichia coli expression system for the A115V mutant of DHODH of T. brucei brucei (sequence identical to that of PDB entry 2B4G 3; we treat this mutant as the wild type in this manuscript) based on pET47b (Merck Millipore) was used, as described previously.11 The gene coding for the C131A mutant was produced by two-step PCR, using subcloning primers (5'- GCTCCCCGGGTCAATGAGCC-3', 5'-CCAGGTACCGCGGCCGCTTA-3') and mutational primers (5'-AATCTGAGCGCGCCGAATGTTCCGGGTAAACC-3', 5'-AACATTC GGCGCGCTCAGATTCAGTTCCAGAA-3'; mutated bases are underlined), which were purchased from Hokkaido System Science (Sapporo, Japan). .. The gene was introduced to the expression vector also as described.11 The proteins were expressed in E. coli strain BL21(DE3) in LB medium containing 30 μg/mL kanamycin (Fujifilm-Wako, Osaka, Japan), with 0.6 mg/mL isopropyl β-D-1-thiogalactopyraonoside (Fujifilm-Wako) at 37°C for 3.5 h. Cells were lysed by 0.5 mg/mL lysozyme (Sigma-Aldrich) in a solution containing 50 mM NaPi (pH 8.0), 500 mM NaCl, 10 mM MgCl2, 5 mM FMN (FujifilmWako), 10 mM imidazole (MP Biomedicals, Santa Ana, CA), 0.025 tablet/mL protease inhibitor cocktail (Roche), 10 μg/mL RNase A (Sigma-Aldrich), and 5 μg/mL DNase I (Sigma-Aldrich), where sonication was also applied. .. After centrifugation, the supernatant was loaded onto a Ni-NTA column (Qiagen) and proteins were eluted with 250 mM imidazole in a solution containing 50 mM NaPi (pH 8.0) and 500 mM NaCl.

    Plasmid Preparation:

    Article Title: Structural Basis of Redox-Dependent Affinities of Dihydroorotate Dehydrogenase for Its Substrates and Products.
    Article Snippet: An Escherichia coli expression system for the A115V mutant of DHODH of T. brucei brucei (sequence identical to that of PDB entry 2B4G 3; we treat this mutant as the wild type in this manuscript) based on pET47b (Merck Millipore) was used, as described previously.11 The gene coding for the C131A mutant was produced by two-step PCR, using subcloning primers (5'- GCTCCCCGGGTCAATGAGCC-3', 5'-CCAGGTACCGCGGCCGCTTA-3') and mutational primers (5'-AATCTGAGCGCGCCGAATGTTCCGGGTAAACC-3', 5'-AACATTC GGCGCGCTCAGATTCAGTTCCAGAA-3'; mutated bases are underlined), which were purchased from Hokkaido System Science (Sapporo, Japan). .. The gene was introduced to the expression vector also as described.11 The proteins were expressed in E. coli strain BL21(DE3) in LB medium containing 30 μg/mL kanamycin (Fujifilm-Wako, Osaka, Japan), with 0.6 mg/mL isopropyl β-D-1-thiogalactopyraonoside (Fujifilm-Wako) at 37°C for 3.5 h. Cells were lysed by 0.5 mg/mL lysozyme (Sigma-Aldrich) in a solution containing 50 mM NaPi (pH 8.0), 500 mM NaCl, 10 mM MgCl2, 5 mM FMN (FujifilmWako), 10 mM imidazole (MP Biomedicals, Santa Ana, CA), 0.025 tablet/mL protease inhibitor cocktail (Roche), 10 μg/mL RNase A (Sigma-Aldrich), and 5 μg/mL DNase I (Sigma-Aldrich), where sonication was also applied. .. After centrifugation, the supernatant was loaded onto a Ni-NTA column (Qiagen) and proteins were eluted with 250 mM imidazole in a solution containing 50 mM NaPi (pH 8.0) and 500 mM NaCl.

    Protease Inhibitor:

    Article Title: Structural Basis of Redox-Dependent Affinities of Dihydroorotate Dehydrogenase for Its Substrates and Products.
    Article Snippet: An Escherichia coli expression system for the A115V mutant of DHODH of T. brucei brucei (sequence identical to that of PDB entry 2B4G 3; we treat this mutant as the wild type in this manuscript) based on pET47b (Merck Millipore) was used, as described previously.11 The gene coding for the C131A mutant was produced by two-step PCR, using subcloning primers (5'- GCTCCCCGGGTCAATGAGCC-3', 5'-CCAGGTACCGCGGCCGCTTA-3') and mutational primers (5'-AATCTGAGCGCGCCGAATGTTCCGGGTAAACC-3', 5'-AACATTC GGCGCGCTCAGATTCAGTTCCAGAA-3'; mutated bases are underlined), which were purchased from Hokkaido System Science (Sapporo, Japan). .. The gene was introduced to the expression vector also as described.11 The proteins were expressed in E. coli strain BL21(DE3) in LB medium containing 30 μg/mL kanamycin (Fujifilm-Wako, Osaka, Japan), with 0.6 mg/mL isopropyl β-D-1-thiogalactopyraonoside (Fujifilm-Wako) at 37°C for 3.5 h. Cells were lysed by 0.5 mg/mL lysozyme (Sigma-Aldrich) in a solution containing 50 mM NaPi (pH 8.0), 500 mM NaCl, 10 mM MgCl2, 5 mM FMN (FujifilmWako), 10 mM imidazole (MP Biomedicals, Santa Ana, CA), 0.025 tablet/mL protease inhibitor cocktail (Roche), 10 μg/mL RNase A (Sigma-Aldrich), and 5 μg/mL DNase I (Sigma-Aldrich), where sonication was also applied. .. After centrifugation, the supernatant was loaded onto a Ni-NTA column (Qiagen) and proteins were eluted with 250 mM imidazole in a solution containing 50 mM NaPi (pH 8.0) and 500 mM NaCl.

    Sonication:

    Article Title: Structural Basis of Redox-Dependent Affinities of Dihydroorotate Dehydrogenase for Its Substrates and Products.
    Article Snippet: An Escherichia coli expression system for the A115V mutant of DHODH of T. brucei brucei (sequence identical to that of PDB entry 2B4G 3; we treat this mutant as the wild type in this manuscript) based on pET47b (Merck Millipore) was used, as described previously.11 The gene coding for the C131A mutant was produced by two-step PCR, using subcloning primers (5'- GCTCCCCGGGTCAATGAGCC-3', 5'-CCAGGTACCGCGGCCGCTTA-3') and mutational primers (5'-AATCTGAGCGCGCCGAATGTTCCGGGTAAACC-3', 5'-AACATTC GGCGCGCTCAGATTCAGTTCCAGAA-3'; mutated bases are underlined), which were purchased from Hokkaido System Science (Sapporo, Japan). .. The gene was introduced to the expression vector also as described.11 The proteins were expressed in E. coli strain BL21(DE3) in LB medium containing 30 μg/mL kanamycin (Fujifilm-Wako, Osaka, Japan), with 0.6 mg/mL isopropyl β-D-1-thiogalactopyraonoside (Fujifilm-Wako) at 37°C for 3.5 h. Cells were lysed by 0.5 mg/mL lysozyme (Sigma-Aldrich) in a solution containing 50 mM NaPi (pH 8.0), 500 mM NaCl, 10 mM MgCl2, 5 mM FMN (FujifilmWako), 10 mM imidazole (MP Biomedicals, Santa Ana, CA), 0.025 tablet/mL protease inhibitor cocktail (Roche), 10 μg/mL RNase A (Sigma-Aldrich), and 5 μg/mL DNase I (Sigma-Aldrich), where sonication was also applied. .. After centrifugation, the supernatant was loaded onto a Ni-NTA column (Qiagen) and proteins were eluted with 250 mM imidazole in a solution containing 50 mM NaPi (pH 8.0) and 500 mM NaCl.



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