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expression of His-tagged protein in insect cells


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#1 Dr Rakesh Kumar

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Posted 18 February 2010 - 05:13 AM

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

Edited by Dr Rakesh Kumar, 18 February 2010 - 05:15 AM.


#2 Vini

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Posted 18 February 2010 - 06:01 AM

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

#3 Dr Rakesh Kumar

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Posted 19 February 2010 - 03:23 AM

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




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