The ReadyPrep protein extraction kit (signal) employs differential solubilization at 4°C in the presence of Triton X-100 to isolate proteins associated with lipid rafts. These proteins are often involved in intracellular membrane trafficking and signaling pathways and include the following: glycosylphosphatidylinositol (GPI)-anchored proteins, caveolin, caveolae (caveolin-associated proteins), acylated tyrosine kinases, heterotrimeric G proteins, and some proteins with transmembrane domains. These proteins are resolubilized in the protein solubilization buffer (PSB), which contains nondetergent sulfobetaine 201 (NDSB 201) along with urea, thiourea, and CHAPS, and which is compatible with a variety of downstream applications, including IEF.
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50 preps, includes 50 ml signal protein extraction buffer 1 (S1), 50 ml signal protein extraction buffer 2 (S2), 25 g protein solubilization buffer (PSB), 30 ml PSB diluent, instruction manual; for 50 extractions of 25–50 mg of cells or tissue