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this protocol utilizes a widely used technique along with reagents and tools that can be acquired commercially to assess endogenous ir5 activation during the early events of stimulation it can easily be applied for probate ir5 biology in other cellular contexts while native page is a widely used technique obtaining clear robust interpretable and reproducible results can be technically challenging using a commercial running buffer and gel system helps to minimize variability and paying attention to details is essential and experience is key to success in obtaining good results this modified native page protocol involves many steps with subtle details that are key to success visual demonstration can show these details that will be beneficial for researchers maintain cal-1 cell culture in a t-75 flask at 37 degrees celsius and 5 carbon dioxide with complete rpmi 1640 medium when ready to stimulate the cells transfer them to a 50 milliliter conical tube and centrifuge them at 200 t
