Heat-Assisted Carbohydrate Antigen Retrieval

Microwave Heat-Assisted Carbohydrate Antigen Retrieval 

Sections (4 xm thick) of formalin-fixed and paraffin-embedded ovarian tissue are mounted on silane-coated slides and air-dried for 24hr at 3°C (Davidson et al., 2000). They are deparaffinized, rehydrated, placed in 0.01 M sodium citrate buffer (pH 6.0), and heated twice for 5 min each in a microwave oven. 2H5 antibody (PharMingen, Becton Dickinson, San Jose, CA) is used at a concentration of 2 p,g/ml to detect sialyl Lex antigen. Staining is performed with labeled avidin-biotin. Negative controls consist of the exclusion of the primary antibody, while positive controls consist of carcinomas that have been shown to be immunoreactive for the antigen in earlier studies. 

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Microwave Heat-Assisted Carbohydrate Antigen Retrieval... 

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