Tlr7 antibody

The TLR7 antibody was purchased from Imgenex Corp., (San Diego, CA, USA), the TLR8 antibody was provided by ProSci Inc. (Poway, CA, USA). COX-2 antibody was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA) and CD34 antibody from Serotec (Duesseldorf, Germany). Isotype control antibodies were purchased by eBioscience (San Diego, CA, USA). Secondary antibodies were Cy3-conjugated AffiniPure Donkey anti-rabbit IgG (Jackson ImmunoResearch Laboratories Inc., Suffolk, UK) and Cy5-conjugated AffiniPure Donkey anti-mouse IgG. The staining was performed on serial cryostat sections of the snap-frozen specimens of pancreatic cancers (UICC II and III) with neighbouring normal pancreas (tumor border) and compared with sections from chronic pancreatitis and normal pancreas. For nuclear counterstaining slides were treated with DAPI (4′,6-Diamidino-2-phenylindoledihydrochlorid) (Sigma-Aldrich, Steinheim, Germany) or haemalaun (Sigma-Aldrich).

Cells derived from normal pancreas, chronic pancreatitis and pancreatic cancer tissues were analyzed on a flow cytometer (Beckman Coulter, Krefeld, Germany) with a software package (Coulter, Epics XL-MCL, System II). TLR7 antibody was purchased from Imgenex, TLR8 was provided by ProSci. CD34-PE antibody, FITC-conjugated anti-rabbit secondary antibody and isotype control antibodies were purchased by Beckman Coulter. For intracellular staining we used IntraPrep kit (Beckman Coulter).