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In some instances, in order to improve the chances of producing a successful antibodies pool, one may choose several target peptides within the same protein sequences and use them as antigen in a co-immunization protocol. Having several epitopes derived from a single protein will increase the chances of obtaining a pool of antibodies that will recognize the target protein. Some background signal increase may be experienced in some instances.
We will synthesize, purify and conjugate each peptide separately. Individual KLH conjugates are then mixed in equimolar ratios and used as immunogen.
An equimolar mixture of 3 KLH-conjugates was used as immunogen in two rabbits. Following co-immunization protocol, antisera were collected and ELISA run. As shown antisera were positive against each individual peptide. The specific response was > 10-fold higher than that of pre-immune sera.