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Example sentences for: ni-

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GST-T7-Cdc14 (RDB 872) and GST-Cdc14 (RDB 1183) were expressed in E. coli BL21(DE3) pLysS cells, and purified on glutathione-agarose beads (Sigma) as described previously [ 33 ] . His6-T7-Net1(1-341) (Net1N, RDB1153) [ 11 ] was expressed in E. coli BL21(DE3) pLysS cells, and purified on Ni-NTA agarose beads (Qiagen) as described previously for p28 Kix1 [ 34 ] . Purification of His6-Plx1, His6-Plx1(N172), and His6-HA-Cdc5 from baculovirus-infected insect cells followed an earlier protocol [ 24 ] . The concentrations of purified proteins were in the ranges of: Net1N~0.
After centrifugation at maximum speed for 15 minutes, the supernatant was incubated with Ni-NTA resin (Qiagen) for 1.5 hour at room temperature.
Recombinant V80R and A61D mutants were expressed in E. Coli BL21 (DE3) pLysS cells (Novagen), and purified by Ni-NTA (Qiagen) and HiTrap SP (Pharmacia) affinity column chromatography followed by ammonium sulfate precipitation as described by Kim et al [ 7 ] .
Upon affinity purification on Ni-NTA resin, HA-Pop1p/His-Myc-Pop2p complexes were incubated with bacterially expressed, phosphorylated Rum1p.
The supernatant fraction was diluted 1:1 with Buffer C (Buffer A containing 0.5% NP-40 and 0.5% deoxycholate, with no SDS) and loaded on to a Ni-NTA spin column (Qiagen).