Imidazol
Imidazol is a chemical compound used in various laboratory applications. It is a heterocyclic aromatic organic compound composed of a five-membered ring containing two nitrogen atoms. Imidazol is often used as a buffer in biochemical and analytical procedures due to its ability to maintain a stable pH range.
Lab products found in correlation
4 protocols using imidazol
Overexpression and Purification of DPYS
ZIKV Methyltransferase Functional Assay
Recombinant Protein Production and Purification
600
of 0.6, followed by induction with 0.4 mM IPTG for 10 h. The cells were then pelleted by centrifugation and stored at −20°C before being lysed.
The cell pellets were resuspended in the selected lysis buffer (20 ml/g cell pellet, 20 mM NaH
2
PO
4
, 500 mM NaCl and 6 M urea, pH 8) (Merck, Darmstadt, Germany) and incubated for 15 min on ice. The suspension was sonicated six times for 20 s and cell debris was removed by centrifugation (13000 g, 30 min, 4°C). The supernatant was passed through 0.45 μm filter before loading on complete His-Tag Purification Column (Roche, Mannheim, Germany). The column was washed extensively with washing buffer (20 mM NaH
2
PO
4
, 500 mM NaCl, pH 8) until the OD
280
of the buffer reached below 0.01. The proteins were then eluted using a column volume of 500 mM imidazol (Sigma-Aldrich), pH 8 containing complete protease inhibitor (Sigma-Aldrich). The eluted fractions were combined and dialyzed against PBS (pH 7.4), and quantified by Bradford total protein assay kit (ZellBio GmbH, Ulm, Germany). LPS contamination was evaluated by the Limulus amoebocyte lysate (LAL) assay (Cambre, USA).
Peroxidase Activity Assay Protocol
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