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Absorption of the His-tagged protein is performed at neutral to slightly alkaline pH to prevent protonation and loss of binding capacity of the weakly basic histidine imidazole groups

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Future studies demonstrated that KEAP1 does not passively sequester NRF2 in the cytosol, but actively functions as a ubiquitin ligase, leading to ubiquitination and proteasomal degradation of NRF2 (Zhang and Hannink, 2003)

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