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Imidazole idz

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Imidazole (IDZ) is a heterocyclic organic compound used as a buffer in biochemical and biological applications. It is a colorless, crystalline solid with a slightly bitter taste. Imidazole serves as a key functional group in various biomolecules, such as the amino acid histidine, and plays a role in pH regulation and catalytic activities in biological systems.

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2 protocols using imidazole idz

1

Protein Expression and Purification

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Restriction endonucleases, DNA polymerases and polynucleotide kinase were purchased from Enzynomics (Daejeon, Korea). Antibodies against hexahistidine (6XHis) epitope or glutathione-S transferase (GST) used for western blotting were from Qiagen (Valencia, CA, USA) and Santa Cruz Biotechnology (Santa Cruz, CA, USA), respectively. Secondary antibodies were from Amersham Biosciences (Piscataway, NJ, USA). Oligonucleotides used in this study were commercially synthesized by Genotech or Macrogen (Daejeon, Korea). All oligonucleotides were gel-purified prior to use. Nucleoside triphosphates were obtained from Sigma-Aldrich (St. Loius, MO, USA), and [γ-32P]ATP (>3000 Ci/mmol) was purchased from Perkin Elmer NEN (Waltham, MA, USA). The pRS plasmids were purchased from New England Biolabs (Beverly, MA, USA). The pET vectors used for protein expression in Escherichia coli were from Novagen (Darmstadt, Germany). Isopropyl β-D-1-thiogalactopyranoside (IPTG) was from ElpisBiotech (Daejeon, Korea). Imidazole (IDZ) was from Acros Organics (Geel, Belgium). The uracil analog, 5-fluoroorotic acid (5-FOA) and proteinase K were obtained from Duchefa Biochemie (Haarlem, Netherland). MMS was obtained from Sigma-Aldrich (St. Loius, MO, USA).
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2

Synthesis and Purification of Protein Complexes

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The oligonucleotides used in this study were synthesized commercially by Bioneer and Genotech (Daejeon, Korea). Oligonucleotides longer than 30 nucleotides (nt) were gel-purified prior to use. The following proteins: Taq DNA polymerases, restriction endonucleases, T4 DNA ligase and polynucleotide kinase were purchased from Enzynomics (Daejeon, Korea). Nucleoside triphosphates were obtained from Sigma-Aldrich (St. Louis, MO, USA). [γ-32P] ATP (>5000 Ci/mmol) was purchased from IZOTOP (Budapest, Hungary). Isopropyl β-D-1-thiogalactopyranoside (IPTG) was from ElpisBiotech, Inc. (Daejeon, Korea) and imidazole (IDZ) was from Acros Organics (Geel, Belgium). The expression vectors (pET21d-MUS81/His-EME1 and pET21d-MUS81/His-EME2) used to prepare the MUS81–EME1/EME2 complexes were kindly provided by Dr Stephen C. West (London Research Institute, UK). In expression vectors, ‘His’ before the name of proteins indicates that hexahistidine tags were fused to the N-terminus of the expressed proteins.
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