Optimization of Glial Fibrillary Acidic Protein Immunoreactivity in Formalin-fixed, Paraffin-Embedded Guinea Pig Brain Sections

摘要

Glial fibrillary acidic protein (GFAP) is an astrocyte-specific intermediate filament whose expression has been shown to be a sensitive marker of toxin-induced brain injury. The present study was designed to employ microwave-assisted antigen retrieval to optimize GFAP immunostaining in formalin-fixed, paraffin-embedded guinea pig brain sections using a variety of commercially available GFAP antibody clones. Of the 7 clones tested for cross- reactivity, following microwave antigen retrieval, mouse clone GA-5 from NeoMarkers was found to be the most responsive, yielding specific GFAP staining in both astrocytic cell bodies and processes. Our observations indicate that different antibody clones must be examined to obtain superior immunolocalization of epitopes uncovered by microwave antigen retrieval. This finding undoubtedly will have important applications in our efforts aimed at determining neuropathological consequences in the guinea pig following exposure to chemical warfare nerve agent.