Table 3.

Purification of 6-His-Tagged AKR1D1 on different HisTrap columns.

	Volume (mL)	mg/mL	Total protein (mg)	pmol/min/μL	nmol/min/mg	Yield % mg	Fold purification	Total activity (pmoles/min)	Yield % activity
Sonicate/Histrap	21	7.42	155.82	44.4	6.0	100	1	931,980	100
Ni Before dialysis	19	0.58	11.02	40.3	70.0	7.07	11.70	766,401	82.23
Co Before dialysis	23	0.53	12.19	32.8	61.9	7.82	10.36	754,566	80.96
Zn Before dialysis	28	0.51	14.28	33.0	64.3	9.16	10.75	922,621	99.00
Ni after dialysis	19	0.51	9.69	28.7	55.9	6.22	9.34	545,681	58.55
Co after dialysis	24	0.39	9.36	24.8	63.7	6.01	10.66	594,510	63.79
Zn after dialysis	29	0.41	11.89	24.6	60.2	7.63	10.07	714,430	76.66
Ni-His Trap plus HiTrap Blue Sepharose	5	0.95	4.75	70.7	74.0	3.05	12.38	353,475	37.93
Co-His Trap plus HiTrap Blue Sepharose	5	1.22	6.10	94.5	77.5	3.91	12.96	472,558	50.70
Zn-His Trap plus HiTrap Blue Sepharose	6	1.37	8.22	107.1	79.1	5.28	13.23	642,553	68.94
10xHis Tag Protein		1.02			80.46

^*Assays measured by fluorimetry.