histag镍柱使用说明书.docVIP

GE Healthcare Instructions 28-4020-89 AD Affinity media Ni Sepharose? 6 Fast Flow CAut Io n! Contains nickel. May produce an allergic reaction. Immobilized metal ion affinity chromatography (IMAC) exploits the interaction between chelated transition metal ions and side-chains of certain amino acids (mainly histidine) on proteins. In general, Ni is the preferred metal ion for purification of histidine-tagged proteins. Ni Sepharose 6 Fast Flow is a BioProcess? IMAC medium that consists of 90 μm beads of highly cross-linked agarose, to which a chelating group has been coupled. This chelating group has then been charged with nickel (Ni2+) ions. Ni Sepharose 6 Fast Flow has low Ni2+ leakage, high protein- binding capacity, and is compatible with a wide range of additives used in protein purification. Its high flow properties make it excellent for scale-up. Ni Sepharose 6 Fast Flow is available in 5, 25, 100, 500 ml, 1L, and 5L bulk packs. Ni Sepharose 6 Fast Flow is also available prepacked in 1 and 5-ml HisTrap? FF, 1 and 5-ml HisTrap FF crude and 20-ml HisPrep? FF 16/10 columns. Contents 1. Product description 3 2. General considerations 5 3. Packing columns 6 4. Packing lab-scale columns 7 5. Packing process-scale columns 8 6. Evaluation of column packing 8 7. Preparation before purification 11 8. Purification procedure 13 9. Optimization 14 10. Troubleshooting 15 11. Regenerating the medium 18 12. Cleaning-in-Place (CIP) 19 13. Storage 20 14. Further information 20 15. Ordering information 20 p. 2 1. Product description Ni Sepharose 6 Fast Flow is highly stable and compatible with a wide range of common additives. This helps to maintain biological activity and increase product yield, while at the same time greatly expanding the range of suitable operating conditions. In addition, the medium is easy to pack and use, and its high flow properties make it excellent for scaling-up. The key characteristics of the medium are listed in Table 1. A variety of compounds