Full length (FL) Nic96 along with C- and N-terminal truncations were constructed and purified as previously described (Fernandez-Martinez et al., 2012 (link); Kim et al., 2018 (link)). Affinity purified and natively eluted full length Nup188 and Nup192 (Fernandez-Martinez et al., 2012 (link)) were incubated with the indicated affinity matrix-bound Nic96 complexes in binding buffer (20 mM HEPES-KOH pH 7.4, 50 mM potassium acetate, 20 mM sodium chloride, 2 mM magnesium chloride, 0.1% Tween-20, 1 mM DTT, 1/250 cOmplete ™ protease inhibitor complex (Sigma-Aldrich)) for 1 h at 4°C. After appropriate washes, the bound complexes were resolved by SDS-PAGE and stained with Coomassie R-250.
Complete protease inhibitor complex
Manufactured by Merck Group
Merck's COmplete™ protease inhibitor complex is a laboratory product designed to inhibit proteases. It provides a broad-spectrum inhibition of serine, cysteine, and metalloproteases.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using complete protease inhibitor complex
1
Purification and Binding of Nup Complexes
2
Purification and Binding of Nup Complexes
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