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After overnight centrifugation, the band containing the nozzles was collected using a gradient fractionator (Labconco Auto Densi-Flow), dialyzed against Tris buffer, and the nozzles were either collected by centrifugation (1 h at 366,000 × g ) or further purified using 30 ml gradients of 10-40% sucrose (wt/wt)
Fractionation Specificity_GST-tagged protein
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