Hi Seb, I'm not aware of the notion (and neither are your reviewers apparently) that a His tag often results in two bands on a lane in SDS page. Why would that be? Extra SDS binding to the positive patch? Just wondering if there's any truth to your statement.
Also, since in this case there seems to be consensus (!) among reviewers regarding the double band, they may have a legitimate point......my 2 cents. Regards, Bert On 10/28/10 7:16 AM, "Sebastiaan Werten" <sebastiaan.wer...@uni-greifswald.de> wrote: Dear all, we have a his-tagged protein that shows a minor accompanying band in SDS-PAGE, just above the main band. According to all other methods available to us the material is homogeneous, the protein has the correct mass in MALDI-TOF, epitopes are recognized, etc. etc. I know that the additional band is a very common artifact with his-tagged proteins, but I was wondering if anyone is aware of a paper that formally describes the phenomenon, as we need to appease a couple of rather bloody-minded referees. Thanks very much for any suggestions, Seb. -- Dr. Sebastiaan Werten Institut für Biochemie Universität Greifswald Felix-Hausdorff-Str. 4 D-17489 Greifswald Germany Tel: +49 38 34 86 44 61 E-mail: sebastiaan.wer...@uni-greifswald.de
