Localization of a 23,000 MW antigen of Cryptosporidium by immunoelectron microscopy

Rabbit antiserum was raised against a 23,000 molecular weight (MW) antigen prepared from Cryptosporidium oocysts by electro-elution from polyacrylamide gels. The antiserum was tested for specificity by immunoblotting against solubilized oocyst preparations. Several antigens including the 23,000 MW antigen were recognized suggesting that it shared common epitopes with higher MW proteins. The antiserum was then used in conjunction with a protein A-colloidal gold conjugate to locate antigenic sites within exogenous and endogenous developmental stages of Cryptosporidium. The pellicles of both sporozoites and merozoites exhibited specific labelling, particularly around their anterior ends. No specific labelling was observed for any other membrane determinants or organelles in these or other life cycle stages.