ReadyPrep™ Protein Extraction Kit (Signal)



Fractionation of Proteins by Cellular Location

Differences in 2-D patterns using ReadyPrep signal (A), membrane I (B), and membrane II (C) kits. Mouse liver samples were extracted using the recommended protocols for each kit. Purified protein (~450 µg) was focused on 17 cm pH 3–10 nonlinear ReadyStrip IPG strips and run on 8–16% PROTEAN II precast gels for the second dimension. Overall spot patterns differ for A, B, and C even though all three kits isolate membrane proteins, indicating each kit isolates different types of membrane proteins.

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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ReadyPrep™ Protein Extraction Kit (Signal), 50 preps
ReadyPrep™ Protein Extraction Kit (Signal), 50 preps

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

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4110149 Instruction Manual, ReadyPrep™ Protein Extraction Kit (Signal), Rev A Click to download