Protein denaturalization for Western blot.

03 October 2011 1 3K Report

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.

Fernando Warleta

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.

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