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After incubation with lysate, beads were washed four times with 3 ml of wash buffer and eluted with 1 ml of elution buffer (5 M of in-house purified SENP EuB protease 51 in wash buffer) at 4 C for 1.5 h with end-over-end rotation

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The cycling rationale relates to receptor sensitivity research: continuous GHSR stimulation may produce receptor desensitization over extended periods in some research models