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文献和实验Purification of 6xHis epitope tagged proteins by Ni-NTA-Agarose His标签蛋白纯化
Low (“Low”)Imidazole Buffer 0.5L 100mM Imidizole 3.4g 5% glycerol 25ml 100% glycerol 50mM Tris-HCl (pH 7.9)50ml 0.5M Tris-HCl (pH 7.9) 0.1% Tween-20 0.5M 100% Tween-20 500mM NaCl50ml 5M NaCl dH2O Fill to 0.5L (start w/ 350ml) High (“High
6xHis-tagged protein purification using Qiagen Ni-NTA Column
)Centrifuge lysate at 10,000xg for 20’ at 40℃to pellet the cellular debris and save supernatant.Add 5μl 2xSB to 5μl Supernatant and store at 200℃for SDS-PAGE analysisBatch purification under Native ConditionsAdd 2.5 ml of the 50% Ni-NTA slurry to 10 ml cleared
One-Step Purification of Recombinant Proteins with the 6xHis Tag and Ni-NTA Resin
The 6xHis/Ni-NTA system is a fast and versatile tool for the affinity purification of recombinant proteins and antigenic peptides. It is based on the high-affinity binding of six consecutive histidine residues (the 6xHis tag) to immobilized
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