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Dimerization of protein


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#1 ventigers

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Posted 13 February 2010 - 06:56 AM

Hello all,

I have purified a His tagged recombinant protein cloned in E.coli BL-21 cells. I suspect that it forms a dimer. I tried running gels under reducing and non-reducing conditions(with and without 2-mercaptoethanol) it gives a single band under reducing and 2 bands under non-reducing conditions. I want to make my point stronger for the sake of publication. can somebody suggest me a method for the same. I came across cross-linking of proteins. Can crosslinking data alone(or with my SDS gels with/without 2-mercap) enough to prove dimerization of my protein of interest.

Venkat.

#2 Vini

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Posted 17 February 2010 - 05:58 AM

Hello all,

I have purified a His tagged recombinant protein cloned in E.coli BL-21 cells. I suspect that it forms a dimer. I tried running gels under reducing and non-reducing conditions(with and without 2-mercaptoethanol) it gives a single band under reducing and 2 bands under non-reducing conditions. I want to make my point stronger for the sake of publication. can somebody suggest me a method for the same. I came across cross-linking of proteins. Can crosslinking data alone(or with my SDS gels with/without 2-mercap) enough to prove dimerization of my protein of interest.

Venkat.



i think u need to go a step further and run a GFC. and correct me if i am wrong, but if its a homodimer why would u see 2 bands??? u should see a band at double the expected molecular weight (or is it that u think that your protein is forming a dimer with another protein??)




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