- Suspend cells by pipetting up and down (Drosophila cells) or
scraping off (mammalian cells).
- Harvest the cells by centrifuging at 2000 rpm for 2 min.
- Discard the medium and suspend the cells in 80 ul extraction buffer.
- Lyse the cells by pipetting up and down.
- Add 20 ul 5x Laemmli solution. Incubate at 100¡C for 5 min.
- Analyse an equivalent of 10 cells per lane by SDS-PAGE.
Extraction buffer:
for example the IP buffer
50 mM Hepes/NaOH pH 7.5, 150 mM NaCl, 1% TritonX100, 10% glycerol,
1.5 mM MgCl2, 1 mM EGTA.
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