His tag protein molecular weight

Georg Wille moqch at mlucom6.urz.uni-halle.de
Fri Oct 23 02:58:46 EST 1998


Ke-xue Huang schrieb in Nachricht <362F6710.8EC39E00 at bioc.rice.edu>...
>Hi, Neuters,
>
>We are trying to detect the molecular weight of Histag expressed protein
>using HPLC gel filtration column, but the proteins come out much smaller
>than that of its immigration on SDS-PAGE. Any suggestion?
>
>Thanks!
>
>Kexue


This ist not unprecedented. In the manual of the QIAexpress system (which
purpose is to construct His-tagged proteins with the subsequent purification
by Ni-chelate chromatography) they describe that 6xHis-tagged
Dihydrofolatereductase DHFR, a protein of ~20 kD molecular weight, runs at
26 kD molecular weight in an SDS-PAGE. They do not say however what the
apparent molecular weight is on HPLC gel filtration.

Georg






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