De-paraffinized and dehydrated sections were treated in a microwave oven at low power for 10 min in 0.01 M sodium citrate buffer (pH 6.0) to retrieve antigen, and inactivated endogenous peroxidase using 3% hydrogen peroxide in methanol for 10 min, and blocked with blocking solution for 1 hour at room temperature. Sections were then incubated with 1∶20 dilution of mouse monoclonal antibody to 8-Hydroxy-2'-deoxyguanosine (ab48508) overnight at 4°C. For detection, a biotinylated anti-mouse and HRP-labeled Streptavidin, and staining with DAB was used.

Immunohistochemical staining of nitrotyrosine (A) and 8-hydroxy-2′-deoxyguanosine (B) was performed on 5 rats randomly chosen from each group (ND: chow diet group, HFD: High fat diet, HFD-QH: High fat diet treated with Qing Huo Yi Hao" (QHYH)). An islet is encircled in each quandrant. Black arrows point to islets of Langerhans (Original magnification ×200). Percentage of immunohistochemial positive staining area in islet out of the area of corresponding islet (C, D) was presented. Values and bars represent means±SEM. * and ** denote PP

ab48508 staining 8-Hydroxy-2'-deoxyguanosine in human colon tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a TE buffer. Samples were incubated with primary antibody (1/50) for 30 mins at 20°C. A HRP-conjugated Goat anti-mouse IgG was used as the secondary antibody.

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