Since you're seeing the MBP band, it sounds as if you're getting some cleavage. 
 If there were no cleave you would only see the fusion and TEV bands on the gel.

It may be that your protein is not stable/soluble without the MBP fusion.  
Different buffer conditions may help if your protein is being lost 

The efficiency of cleavage may also be aided by different buffer conditions.  
Since you didn't report the buffer you're using, it's impossible to judge if it 
is appropriate for maximum TEV activity.


Sent from my iPhone

On Nov 4, 2012, at 10:24 AM, "rana ibd" 
<<>> wrote:

Dear CCP4
     I am having a problem with cleaving my fusion protein and I would be 
grateful if you advice me regarding this situation,  I have an MBP-DHBx fusion 
protein and I am trying to cleave it using TEV protease, I have tried different 
ratios and different temperatures  with different incubation time but still it 
will not cleave, all I observe on the gel is the bands of the fusion protein 
which is 59kDa and the MBP which is 42kDa and the TEV protease which is 27kDa 
and no sign of the DHBx which is 17kDa,I have also checked the sequence if 
there was any problem but I could not find anything unusual the sequence was 
fine , so if you have any suggestions regarding this situation I will be 
Best Regards

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