2 replies to this topic

[Dr Rakesh Kumar]

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.

[Vini]

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

[Dr Rakesh Kumar]

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