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Clusterin is an enigmatic glycoprotein with a nearly ubiquitous tissue distribution. It plays important roles in protein homeostasis/proteostasis, inhibition of cell death pathways, and modulation of pro-survival signaling and transcriptional networks.
Clusterin Purification Protocol Using Affinity Chromatography
HEK293 cells stably transfected with clusterin were seeded in 10cm plates and allowed to grow to confluence (typically 2-3 days). Cells were then trypsinized and seeded into 75cc flasks where they were allowed to grow for 7-10 days. The supernatant was harvested from these flasks and centrifuged for 10 minutes at 1000 g to pellet any detached cells or debris, then filtered through a 0.22-micron filter and kept on ice until applied to the affinity column.
Fig.1 Flow chart of clusterin purification protocol using affinity chromatography. (Creative Biolabs)
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