Protein hispurtm ni nta purification kit

The Escherichia coli strains DH5α and BL21(DE3) pLysS, as well as expression vector pET-32a (+), were purchased from Novagen (Darmstadt, Hesse, Germany). Restriction enzyme Kpn I, EcoR I and Mlu I, T4 DNA ligase, protein markers, protein HisPur^TM Ni-NTA purification kit, Fetal Bovine serum (FBS), penicillin, streptomycin, and cRNA mMESSAGE mMACHINE In Vitro Transcription Kit were purchased from Thermo Scientific (Waltham, MA, USA). enterokinase was purchased from New England Biolabs, Inc. (Ipswich, MA, USA). Vydac C18 columns (5 μm, 4.6 mm × 250 mm, 10 μm, 22 mm × 250 mm) were purchased from Grace (Deerfield, IL, USA). Acetonitrile (ACN, gradient grade for HPLC), Acetylcholine chloride (ACh) and other chemical reagents were all of analytical grade and purchased from Sigma-Aldrich (St. Louis, MO, USA).

The Escherichia coli strains DH5α and BL21(DE3) pLysS, as well as the expression vector pET-32a containing the Trx tag, were purchased from Novagen (Darmstadt, Germany); pIVEX 2.4d was purchased from Roche (Basel, Switzerland); and pET-SUMO-28a was constructed and conserved in our lab. The RTS^TM 100 Escherichia coli Disulfate Kit was purchased from Biotechrabbit GmbH (Berlin, Germany). Restriction enzymes Kpn I, EcoR I, Not I, and Mlu I, T4 DNA ligase, protein markers, protein HisPur^TM Ni-NTA Purification Kit, HAM’S/F12 medium, Fetal Bovine serum (FBS), penicillin, and streptomycin were purchased from Thermo Scientific (Waltham, MA, USA). Enterokinase was purchased from New England Biolabs, Inc. (Ipswich, MA, USA). Vydac C18 218TP54 columns (5 μm, 4.6 mm × 250 mm, 10 μm, 22 mm× 250 mm) were purchased from Grace (Deerfield, IL, USA). Acetonitrile (ACN, gradient grade for HPLC), and other chemical reagents were all of analytical grade and purchased from Sigma-Aldrich (St. Louis, MO, USA).