Q. About Mouse Anti-His Tag Ab!

[Miss ELLA CHAN]

   Dear all,

   I have transformed the pET16b vector with a insert in it in BL21
   and performed protein expression. pET16b vector is one of the
   expressing vector from the pET system which allows the expressed
   target protein to be linked with polyHis so that it will be easier
   for purification by using His-tagged Affinity Column.
 
   Now, I would like to check my expressed target protein by doing Western
   Blot using the mouse anti-His tag HRP conjungated antibody. However, it 
   gave me no bands that was cooresponding to the expressed one which I have
   been seen on SDS-PAGE Gel.

   So does it mean that the expressed one is not polyHis linked 
   or the antibody is not suitable for testing BL21 expressed protein??
   Is there any other way to confirm the expressed protein is the one I want 
   ( other from checking the molecular size ) ??

   Thanks so much for your valuble suggestion!!

   Regards,

   Ella Chan Wai Ching

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