problems with electrophoresis

[buthercup]

Hi every one
I hope some people could help me. I’m quite disappointing and perplexed, because after performing a protein extraction from bacteria, with a typical 2D buffer (Tris, Urea, Thiourea, CHAPS), I quatify the extraction and obtain about 4 ug/ul, quite a big concentration. But in the moment that I perform a 1D-SDS PAGE, I don’t see any band on the gel. Of course I mix the desired volume of my protein extraction with the Sample Buffer for monodimensional gels, in the correct proprtion (final concentration of Sample Buffer = 1X)
Do any of you have an idea of where is the problem?
Thank you in advance

[JoB]

2 questions:

1) How much protein do you load per well?
2) What stain do you use to visualize?

[Author]

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