His-tag inhibits anti-myc-antibody

wulf-dirk.leuschner at tuebingen.mpg.de wulf-dirk.leuschner at tuebingen.mpg.de
Thu Jan 29 21:37:02 EST 1998


Hi there,

we had a couple of difficulties lately using constructs with a
His6-tag fused to other epitope-tags (at the N-terminus of various
proteins): using either a His6-HA-Tag or a His6-myc6-tag we only got
very weak staining in W.Blots and dot blots (using either the
12CA5-anti-HA antibody or the 9E10-anti-myc antibody). Has anybody had
similar problems?
We believe that it is the His6-tag which interacts somehow with the
tags (the HA and myc-tag both have a couple of negatively charged aa
residues ...). At the moment we have no idea how to overcome this
inhibition/interaction. Does anybody have an idea what we could do?
(Adding mM-amounts of Ni2+ does not change a lot...)

WDL




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