Cell lysis and Lowry assay for 2D electrophoresis

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      samkh918
      Participant

      Hello,

      I want to perform a 2D electrophoresis on my neuronal cells and wanted to get your feedback on some of my questions:

      1)Can I use my 2D sample buffer:

      8M Urea,
      2% w/v CHAPS
      50mM DTT
      0.2% w/v100X Bio-Lyte ampholyte
      0.002% w/v BPB

      for the cell lysis step as well? I am going to try both sonication and grinding techniques for cell lysis

      2)Can I use Lowry assay if I use this 2D sample buffer for cell lysis. I know that Lowry assay is not reducing agent compatible, but the DTT concentration in this 2D buffer is 50mM. Do you think it will interfere with the Lowry assay?

      3)I assume I can’t use Bradford assay here since I am using a detergent, CHAPS.

      Thanks a lot for your help.

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