problems with electrophoresis

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    • #10902
      buthercup
      Participant

      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

    • #89234
      JoB
      Participant

      2 questions:

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

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