Hi!
At 10Kd,. you probable are masking your protein with unincorporated
label, not the Heme protein, I think (unless you are running very high
percentage gels). Cant you do your TnT and then IP to clean it before
running it out on a gel to see if its there? I had to do that with a
11Kda piece of protein to see it.
Good luck!
John
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John Strasswimmer, MD,PhD Candidate | Phone (617) 636 8396
Tufts - New England Medical Center | Fax (617) 636 6190
Box 166 | Email: jstrassw at opal.tufts.edu
Boston, MA 02111 USA |
HTTP://WWW.healthsci.tufts.edu/microbiology/strass/JSpage.HTML
*** Co-Author of "HyperBug" Microbiology Teaching Software ***
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On 22 Oct 1996, D.L. Roberts wrote:
> I have been trying to produce a 35-S labelled protein using Promega's
> rabbit reticulocyte lysate kit and am having problems as one of the
> fragments of the active enzyme is approximately 10kD and it is being
> masked by Haeme proteins which run at about the same position. Has
> anyone any suggestions on removing the haeme protein or know of a method
> of preventing it picking up the radiolabel?
>
> Thanks in advance.
>
> Darren Roberts
>
> MRC Toxicology Unit,
> University of Leicester.
>
> DLR2 at le.ac.uk
>
>
>