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文献和实验One-Step Purification of Recombinant Proteins with the 6xHis Tag and Ni-NTA Resin
nickel ions, giving a highly selective interaction that allows purification of tagged proteins or protein complexes from 95% homogeneity in just one step (1 ,2 ). The tight association between the tag and the Ni-NTA resin allows contaminants
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
Extraction of DNA from Agarose Gels
, to destabilize the agarose gel. The DNA is subsequently bound to a substrate (e.g., an anionic resin) and washed to remove impurities prior to elution of the DNA from the substrate. Other methods include hot phenol extraction of the DNA from the gel. The use
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