蛋白质分离纯化与定性定量分析.pptVIP

Ion-Exchange chromatography Ion-Exchange chromatography Size-exclusion chromatography Size-exclusion chromatography Affinity chromatography Affinity chromatography Commonly used affinity columns: Ni2+ ? binds to poly Histines (example 6xHis) Specific antibodies (anti-Flag tag) glutathione ? binds to GST Protein A or G ? binds antibodies Affinity chromatography Possible elution strategies: pH Ion strengh Denature Competitor ligand or analog Ni-NTA columns The high affinity of the Ni-NTA resins for 6xHis-tagged proteins or peptides is due to: 1- the strength with which these ions are held to t