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expression of His-tagged protein in insect cells - (Feb/18/2010 )

hi every body
I have expressed his-tagged protein which is of about 53KDa. But it is showing about 65KDa size on SDS-PAGE. Recombinant protein is also reacting with specific polyclonal sera. Why there is size descrepency i dont know. May be Histidine 6X tag influences the mobility of protein in SDS-PAGE. Can any body help me out.

thanks

-Dr Rakesh Kumar-

Dr Rakesh Kumar on Feb 18 2010, 07:43 PM said:

hi every body
I have expressed his-tagged protein which is of about 53KDa. But it is showing about 65KDa size on SDS-PAGE. Recombinant protein is also reacting with specific polyclonal sera. Why there is size descrepency i dont know. May be Histidine 6X tag influences the mobility of protein in SDS-PAGE. Can any body help me out.

thanks



No, his tag will not cause this change in mobility. This topic on anomalous mobility of proteins in SDS-PAGE has been discussed previously in this sub-forum. It could be due to post-translational modification of ur protein

-DRN-

DRN on Feb 18 2010, 08:31 PM said:

Dr Rakesh Kumar on Feb 18 2010, 07:43 PM said:

hi every body
I have expressed his-tagged protein which is of about 53KDa. But it is showing about 65KDa size on SDS-PAGE. Recombinant protein is also reacting with specific polyclonal sera. Why there is size descrepency i dont know. May be Histidine 6X tag influences the mobility of protein in SDS-PAGE. Can any body help me out.

thanks



No, his tag will not cause this change in mobility. This topic on anomalous mobility of proteins in SDS-PAGE has been discussed previously in this sub-forum. It could be due to post-translational modification of ur protein



thanks .. i will work on your suggestions

-Dr Rakesh Kumar-