https://eln.excellgen.com/index.php?action=history&feed=atom&title=Ni-Coated_Plate 2026-10-19T05:38:29Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=Ni-Coated_Plate&diff=20010&oldid=prev 2019-06-28T02:00:57Z

<p>Created page with &quot;[https://bmcbiotechnol.biomedcentral.com/articles/10.1186/1472-6750-8-16 https://bmcbiotechnol.biomedcentral.com/articles/10.1186/1472-6750-8-16] Preparation of Ni2+-NTA coat...&quot;</p> <p><b>New page</b></p><div>[https://bmcbiotechnol.biomedcentral.com/articles/10.1186/1472-6750-8-16 https://bmcbiotechnol.biomedcentral.com/articles/10.1186/1472-6750-8-16]<br /> <br /> Preparation of Ni2+-NTA coated microtiter plates The Ni2+-NTA coated microtiter plate was prepared as described previously&amp;nbsp; except an ordinary ELISA carboxylated polystyrene microplate (polystyrene high BIND microplate, Corning life science, USA) was used instead of a plate coated with maleic anhydride (Pierce, USA) as the later is 20 times more expensive. <br /> <br /> Briefly, a microtiter plate was coated with N,N-bis [carboxymethy]lysine (Sigma Aldrich, USA) overnight and washed 3 times with 0.05% Tween 20 before blocking with 3% BSA in 50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 0.05% Tween 20. The coated plate was washed with a series of buffers including 1) 50 mM Tris-HCl, pH 7.5, 500 mM imidazole, 0.05% Tween 20 2) 0.05% Tween 20 3) 100 mM EDTA, pH 8.0 and 4) 0.05% Tween before charging with 10 mM NiSO4 for 20 minutes at room temperature. After washing with 0.05% Tween 20 and then with 50 mM Tris HCl, 500 mM NaCl, pH 7.5, the plate was then ready to be used to capture (His)6-containing proteins.<br /> [[Category:Protocols]]</div>

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