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. 2006 Jun 2;39(1):151–156. doi: 10.1016/j.ijbiomac.2006.05.010

Fig. 1.

Fig. 1

Metal chelate affinity chromatography. (A) Purificaton yield. Percentage of IC1-His protein in the cellular supernatant before (black bar) and after (white bar) loading into the chelating column, both for non-diluted (1) and 1:10 diluted (2) samples. The OD at 490 nm resulting from ELISA assays were normalized to that of non-diluted initial cell supernatant. (B) Efficiency of protein elution. OD at 490 nm from an IC1 ELISA assay of elution fractions from the HiTrap-Chelating column (see Section 2): (1) fraction 2 imidazol 250 mM; (2) fraction 3 imidazol 250 mM; (3) fraction 4 imidazol 250 mM; (4) fraction 5 imidazol 250 mM; (5) fraction 1 imidazol 500 mM; (6) fraction 2 imidazol 500 mM; (7) fraction 3 imidazol 500 mM; (8) fraction 4 500 mM. Protein samples were diluted 1:10 prior to the ELISA test. (C) Purity of column elution fractions containing IC1-His. A 10% SDS-PAGE of cellular supernatant before (b) and after (a) loading into the HiTrap-Chelating column and of fractions indicated in panel B. Inset on the right shows western-blot of IC1 immunoprecipitates from transfected (+) and untransfected (−) 293T cell supernatants. Size and migration of molecular weight markers are indicated.