recombinant protein form - (Feb/14/2013 )

Hi all
how to identify how my recombinant proetin is expressed in E. coli i,e. as inclusion body or as free form before going for use of NiNTA puriifcation so as to help in deciding the approprite method of puriifcation under native or under denaturing condition.

please give a good reliablesource also so as help in citing in publication


thanks

This handbook might be helpful- it's discussed in chapter 6 and several other parts of the book: http://wolfson.huji.ac.il/purification/PDF/Tag_Protein_Purification/AMERSHAM_TheRecombProtHandbook.pdf

Another useful resource for solubility studies and purification: http://www.embl.de/pepcore/pepcore_services/protein_purification/extraction_clarification/invitro_denaturation_refolding/

We check in miniscale culture (5-10 ml) first then go for large scale purification. Just spin the culture, lyse cells and spin again to separate pellet and supernatant. Load both on gel you'll get to know where is your protein going.