Hi, I have to do an hypotetical research on a new gene, associated with hair growth, and make a polyclonal antibody. The only thing I am not sure about is which method to use for the preparation of antigen samples.
Thanks for the help
Hi Selene
you have two options : to find antigenic peptides in your protein using some bioinformatic tools and synthetise them in vitro
http://www.bioinformatics.nl/cgi-bin/emboss/antigenic
To express in E.coli an purify one histidine tagged hydrophilic fragment of your protein 300-400bp (100--150 aa) in length , and use it to immunise guinea pig or rabbit, good luck
If you prefer to make synthetic peptides I'd recommend you add a cys residue at N or C terminus. This will make the production of the immunogens much easier, as you will need to conjugate peptides to KLH (for immunisation) and BSA (for screening of the sera). The cys residues allow easy coupling to maleimide derivatives of the above two proteins. mal-BSA and mal-KLH are commercially available if you don't know how to make them yourself.
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