M o m immunodetection peroxidase kit

Surgical specimens were separated into two samples for histopathological and immunohistological analyses of the α v β 3 integrin expression level. A portion of the samples was fixed in formaldehyde, embedded in paraffin and cut into 7 μm slices for hematoxylin and eosin (H.E.) staining of the orthotopic tumors, residual fluorescent tissues and non-fluorescent control tissues. Histological examination was performed under an Olympus BX41 Laboratory Microscope. Any tumor foci identified in the residual fluorescent tissues were measured using the Image J 1.46r software. Other samples were frozen in isopentane and underwent shock freezing in liquid nitrogen. These samples were cut into 8 μm slices (microcryotome) and stored at -80 °C until fixation and staining. After the frozen slices were thawed at room temperature, they were fixed in acetone. Immunostaining with a mouse anti-human integrin α v β 3 monoclonal antibody (clone MAB1976, 1:500; Millipore) was performed on the acetonefixed cryosections of the orthotopic tumours using the M.O.M. immunodetection (peroxidase) Kit (Vector laboratories, Inc., Burlingame, CA). Staining was performed using liquid DAB and a substrate chromogen system (Dako, North America, Inc., CA). The nuclei were counterstained with hematoxylin.