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Hello, thank you for your protocol. I'm interested in peptide...
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vu quang hung Oct 20, 2020
Hello, thank you for your protocol. I'm interested in peptide 2 kDa. I have tried your protocol: 18% glycine - SDS- PAGE, 25 A for 3,5 hours. I use ruler 10- 250 kDa. The green band is 10kDa. And this is the results. Nearby the green band is 2 lanes of bromophenol (0,6 kDa as you wrote). 
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jose david rosales Answered Oct 21, 2020
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Good morning, in the gel I see that the green band is very close to the end of the gel; which could indicate that the low molecular weight band came out of the gel during the electrophoretic run.
In the gel that I show the 10 kDa band is in the middle of the gel. Please check the acrylamide concentration of your gel.
You can use 20% acrylamide and 20 minutes of pre-run.
It does not change the urea concentration.
The amperage can be 30 mAmp, but take care of the temperature, that the buffer does not overheat, a cold room is useful.
In the gel that I show the 10 kDa band is in the middle of the gel. Please check the acrylamide concentration of your gel.
You can use 20% acrylamide and 20 minutes of pre-run.
It does not change the urea concentration.
The amperage can be 30 mAmp, but take care of the temperature, that the buffer does not overheat, a cold room is useful.
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