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Gel running
10% Bio-rad pre-casting gel, 200volts all the way, 40-45min.
or
Hand-casting gel, 4% stacking, 12% resolving, 80volts when in stacking(10-15 min), change to 200volts when you see bromophenol blue in resolving gel (35-40 min). You can get the band as straight as drawing with a ruler.

Transfer
200 volts, 2 hr, 4 degree.
or 80 volts, overnight, 4 degree.

I used to do a lot of WB for a protein at 83kD, if you need, I can send you my protocol.

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3Â¥2015-04-25 01:43:56
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