Determining Optimal Microwave Antigen Retrieval Conditions for Microtubule-Associated Protein 2 Immunohistochemistry in the Guinea Pig Brain
Technical rept. Jan-May 2002
ARMY MEDICAL RESEARCH INST OF CHEMICAL DEFENSE ABERDEEN PROVING GROUND MD
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The present study examined the efficacy of microwave pretreatment on microtubule-associated protein 2 MAP-2 immunoreactivity in paraffin-embedded sections of formalin-fixed guinea pig brains using different MAP-2 monoclonal antibodies. Brain sections were boiled in sodium citrate, citric acid, Tris hydrochloride, and EDTA solutions with pH values of 2, 4, 6, and 8 in a microwave prior to MAP-2 immunohistochemical staining. Specific MAP-2 immunoreactivity was observed in brain regions when NeoMarkers MAP-2 antibody clone AP-18 was used in conjunction with citric acid buffer of pH 6.0 as an antigen retrieval solution. No immunoreactivity of MAP-2 was observed in negative control sections. The results suggest that a lO-min boiling in citric acid solution at pH 6.0 is the optimal microwave-assisted AR method for immunolabeling MAP-2 in formalin-fixed, paraffin-processed guinea pig brain samples using NeoMarkers MAP-2 monoclonal antibody AP-18. This undoubtedly will have important applications in our efforts to conduct retrospective studies on archival guinea pig brain paraffin blocks, ultimately relaxing the use of additional animals to evaluate changes in MAP-2 expression between chemical warfare nerve agent-treated and control samples.
- Anatomy and Physiology
- Medicine and Medical Research