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2-D Gel Problems with IEF

2-D Gel Problems with IEF - Proteomics Forum

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  #1  
Old 11-28-2006, 09:19 PM
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Question 2-D Gel Problems with IEF



Hey guys,
I am having problems with my IEF (isoelectric focusing). It seems the second dimension is fine, but the IEF is messing up somehow. I think it may be the voltages.

Does anyone have any recommendations for the voltages?

thanks
Kiki
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Old 03-26-2007, 11:02 PM
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Default Re: 2-D Gel Problems with IEF

Using Biorad Ready Gels I use the following settings:

100V 60 min
250V 60 min
500V 60 min

25mA Max
10W

I run the gels using NaOH in the inner chamber and Phosphoric acid in the outer chamber. Works well for me.
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Old 05-11-2007, 06:26 PM
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Default Re: 2-D Gel Problems with IEF

what exactly is ur problem ...? make it little more elobarate...
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Old 05-25-2007, 06:34 AM
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Default Re: 2-D Gel Problems with IEF

Using the bio-rad IPG strip,for 17cm,passive absorption,i use the following step:250V 30 min
1000V 1h
10000V 5h
10000 60000Vh
500V anytime
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Old 05-29-2007, 01:27 AM
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Default Re: 2-D Gel Problems with IEF

Hi
Whts the programme you gave for IEF and wht type of fluctations you notice. write in detail
regards
SIvaramaiah
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Old 06-04-2007, 12:01 AM
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Default Re: 2-D Gel Problems with IEF

As condições de corrida irão depender da faixa de pH e do tamanho da strip.
Em que faixa você esta utilizando???
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Old 11-10-2008, 07:17 PM
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Default Re: 2-D Gel Problems with IEF

Hi here is what im using, but mine isnt working either.
250V 15 minutes
4000V, rapid ramp, totaling 20000vh
500V hold step.
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