Envision dab detection kit

Four μm thick sections of the formaldehyde fixed and paraffin embedded BxPC3 and PancTu-1 xenograft tumors were exposed to the rabbit monoclonal Anti-Prostaglandin E Receptor EP2/PTGER2 antibody (Abcam, USA; cat.no.: ab167171) at a dilution of 1:1000 following deparaffination, rehydration and antigen retrieval (pH 6.0) as usual. A HRP-labeled anti-rabbit polymer antibody (Mach2, BioCare Medical, USA, cat.no. RHRP520) and the Envision DAB detection kit (DAKO-Agilent Technologies, USA) were used for detection and visualization of the specific antibody binding, followed by hematoxylin counterstaining. Microscopic images were taken by a Leica DM2500 research microscope equipped a DFC495 digital camera and the LAS imaging tool kit.

For the demonstration of APN/CD13 expression, 4-5 μm thick sections of the formaldehyde fixed and paraffin embedded He/De and Ne/De tumors were exposed to the mouse anti-rat CD13 (Santa Cruz Biotechnology) at a dilution of 1:50 following deparaffination, rehydration and antigen retrieval (at pH 6.0). A HRP-labelled rabbit anti-mouse polymer antibody (Abcam, UK) and the Envision DAB detection kit (DAKO-Agilent Technologies, USA) were used for visualization of the specific antibody binding, followed by hematoxylin counterstaining. Photos were taken with Nikon Eclipse E800 (Nikon Corporation, Tokyo) microscope.