detection diagnosis quantitation staging and classification of normal and activated ras p21s in normal tissues malignant and premalignant lesions application_number US 7117587 A description to produce murine monoclonal antibodies ras 8 10 and 11 balb c x c57b1 6 mice were immunized intraperitoneally i p with 100 micrograms u03bcg of a recombinant ras p21 protein produced in bacteria the protein used for immunization was of the harvey ras family and contained the amino acid arginine at position 12 instead of the normal amino acid glycine since the protein contained a mutation at position 12 the protein used for immunization is considered to be of the oncogenic or activated form the first and second inoculations consisted of a mixture of complete freunds adjuvant and 100 u03bcg of the activated p21 subsequent inoculations given at two week intervals consisted only of the activated p21 protein ras 8 was produced in fusion 351 mouse 3103 ras 10 was produced in fusion 352 mouse 3104 and ras 11 was produced in fusion 353 mouse 3105 mouse serum was collected and evaluated by elisa for the presence of anti p21 antibody mice were selected for fusion based on the degree of serum reactivity to the immunogen three days before immune spleen cells were used for fusion.