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protein purification before antibody production
I have a question on the topic which is in the title and I hope that somebody could help me.
I would like to produce antibody and as a antigen use whole protein (not some peptide) and my protein is quite big- 250 kDa. What kind of method of purification is the best in such situation?
And the second question is if it is ok for immunization if the protein contain c-Myc tag?
thank you in advance for ever answer.
Re: protein purification before antibody production
Hola,It doesn´t matter if you inmunize with a taggeg ptotein, moreover you need it to purify. The problem will be if the pure protein will be soluble or not, and probably second option (by the size) will be more probable. and immunizing with denatured protein could generate antibodies against hided epitopes in the nature protein, but for policlonal it probably recognizes other epitopes of the nature protein, and for monoclonals you have to assay each clon against the natural protein to choose the best. Some ocassions I had to extract stainde protein of a band of preparative PAGE to immunize rabbits and it runs. Buena suerte
|antibody , production , protein , purification|
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