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3T3-L1 Plasma Membrane Isolation

28 bytes added, 21:44, 21 January 2011

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#Collect supernatant and spin in TLA 100.3 rotor at 19,000 rpm for 20 minutes at 4 degress (Tube = Beckman #349622)

#(Optional) Wash pellet in 1.0 mL HES and spin in TLA 100.3 rotor at 19,000 rpm for 20 minutes at 4 degress (Tube = Beckman #349622)

#(Optional) To further purify PM, resuspend in 0.6 mL HES and repeat steps #4 and #6

#Resuspend pellet in 1.0 mL HES and homogenize with dounce for 10 strokes

#Layer the resuspended pellet on top of 10 mL 40% sucrose/HES in a 13.5 mL ultracentrifuge tube (tube = Beckman #331372)

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