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In vitro translation problems

jstrassw at OPAL.TUFTS.EDU jstrassw at OPAL.TUFTS.EDU
Tue Oct 22 15:43:11 EST 1996

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 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                   |                              
     *** 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

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