Muc Glycoprotein Antibodies

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Publication protocol

Formalin-fixed paraffin embedded tissue blocks of the cases were retrieved and five sections prepared, one for H&E and four, for the IHC stain. For immunostaining, sections were deparaffinized in xylene and rehydrated in alcohol. Endogenous peroxidase activity was inhibited by incubating the sections in 3% H2O2 for 20 minutes. For antigen retrieval, specimens were boiled in a citrate solution (pH=6) for 20 minutes. All the sections were incubated in goat serum for 20 minutes, after which slides were incubated overnight with primary monoclonal antibodies at 4˚C. Sections were rinsed and incubated for 30 minutes with secondary antibody, horseradish peroxidase conjugated anti-mouse and rabbit immunoglobulin. Then, sections were rinsed and incubated with DAB for 5-7 minutes and counterstained with hematoxylin. Table 1 shows the characteristics of each antibody used for IHC.

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