I am wondering if extracellular protein are also taken out when we extract protein directly from a dish (PBS-washed cells). Many protocols appear to detach cells with EDTA and then extensively boil the remaining material with SDS to extract them.
Attila Lehotzky
it depends on the extraction buffer composition. RIPA likely solubilizes them, for example.
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Gaetan Mayer
After detaching the cells, scrapping the dish with hot Laemmli sample buffer works to solubilize extracellular proteins strongly attached to the plastic. Using small volumes allow you then to load the samples on SDS-PAGE gels
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