2 replies to this topic

[xabiarias]

Hi all,

I have succesfully cloned a gen in pET28a through BamHI and XhoI sites. My goal is to purify the corresponding protein and inoculate it into rabbit for immunization. The point is that as it has been cloned through BamHI site the recombinant protein will have the His-tag plus the T7-tag (MASMTGGQQMG).

I am afraid that the rabbit could produce undesirable antibodies through this N-terminal region of the recombinant protein. Does anybody knows whether the T7-tag is immunogenic?

Thanks in advance,

[Chelo]

Dear Xabiarias,

the T7-tag might develop an antibody response. However, if you are producing polyclonal antibodies, you don´t need to worry about that since you will have protein specific antibodies too.
Regards.

[xabiarias]

That's right Chelo but afterwards, if I perform a immunochromatography purification step with the recombinant protein coupled, I will certainly purify those antibodies too...