Hello,
I have a problem during protein denaturalization for SDS-PAGE electrophoresis. I am adding 100 uL of 1x sample buffer directly over 1x10E5 tumor cells, heatting 5' at 90 ºC and cooled. When I am going to charge proteins into electrophoreisi gel, sample looks like a mucus and is imposible to load it.
Can anyone help me?, thanks a lot.
Thank you Gael for your answer.
I will tray today to shear DNA through an insulin syringe 5 times, because we don´t have a sonicator and to avoid boiling and cool on ice.
Yesterday we tried it. In fact, on first samples we made a lot of foam, but on last samples we done it very carefully avoiding foam and mucus disappeared! Thank you for your appreciation about sample buffer concentration, I've read about it and we conclude as same as you propouse; 2x sample buffer directly on cell samples and 1x on Molecular Weight and on comercial protein extract (positive controls).
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