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Cells were sonicated five times for 30 seconds each with 1 minute cooling between each cycle, then centrifuged at 15,000 rpm for 10 minutes at 4°C. 0.5 ml of Talon resin (Cat 635501, Clontech) equilibrated with lysis buffer was then added to the supernatant and gently mixed at 4°C for 2 hours.
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Do 20 crunches and 30 jumping jacks with a two minute cool down between them.
9. 3 - 5 minutes cool-down.
(30 - 40 minutes) Cool the cakes.
Following a 10-minute baseline, subjects received 30 minutes head cooling followed by 30 minutes head and neck cooling.
Each of these exposures lasted for 10 minutes plus approximately 10 minutes for cooling the body with a cold shower and a rehydration between the trials.
Allow a minute for cooling down.
40 minutes, plus cooling time.
Total time: 20 minutes, plus cooling time.
Prep Time: 20 minutes, plus cooling time.
Prep Time: 25 minutes, plus cooling time.
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