Table 1. Recommended electrophoresis conditions for IPG gels on dedicated IEF systems

Table

Step	IPG length (cm)	Voltage	Duration (hr:min)	Volt-hours	Gradient type
1	7, 11, 13, 18, 24	rehydration	12:00^a	-	-
2	7	500	0:30	250	step and hold
2	11, 13, 18, 24	200	1:00	500	step and hold
3	7	8000^b	1:00	8000	step and hold
3	11, 13	8000^b	2:00	16,000	step and hold
3	18	8000^b	4:00	32,000	step and hold
3	24	8000^b	6:25-11:25	50,000-90,000	step and hold
^aThe total rehydration time can be varied for convenience, but it must be >6 hours.
^bDepending on the sample, this voltage may not be reached within the suggested step duration. For this reason, it is best to program the steps based on volt-hours rather than time.