Anti-His tag antibodies

syan syan at med.unc.edu
Thu Sep 25 13:45:32 EST 1997


Has anyone out there had any success using anti-His tag antibodies for
western blots?  We have tried the anti-tetra and penta His MAbs from
Qiagen for western blots of two different proteins which have C-terminal
6xHis tags, and had no success.  We are pretty sure the His-tag is
intact on the proteins, given that the proteins were purified with
Ni-NTA resin immediately prior to the western blot analysis.  The Qiagen
tech person suggested trying different blocking agents (BSA, powdered
milk, and casein).  We tried these, with no luck. Has anyone (not
associated with Qiagen) compared Qiagen's anti-His Abs to those from
other companies?




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