Decreased D2-40 immunoreactivity in stored paraffin sections and methods for preserving it

Abstract

D2-40, a monoclonal antibody against podoplanin, is a selective marker of lymphatic endothelium and is widely used for research on and diagnosis of pathology of lymphatic vessels. We examined the relation between the duration of tissue section storage and changes in immunostaining by D2-40 antibody; we evaluated also the effects of preservation methods on changes in immunostaining during storage. Staining by D2-40 was attenuated by long-term preservation of scalp skin and lymph node sections at room temperature. The attenuation of D2-40 staining in stored sections was improved by preservation at low temperature, i.e., 4° or - 30° C. We investigated also the immunostaining of preserved tissue sections using NZ-1 and Lyve-1, which are antibodies against lymphatic endothelium markers. Staining by NZ-1 or Lyve-1 antibody was detected clearly in sections that had been stored for 16 weeks. Our study suggests that either long-term storage of D2-40 immunostained tissue sections should be avoided or the section should be preserved at low temperature.

Bone morphogenetic protein-4 and bone morphogenetic protein receptors expressions in the adult rat eye

“…They were embedded in routinely paraffin wax, serially sectioned at 4 μm and mounted on glass slides, and these sections were preserved at -30°C until use to prevent the attenuation of immunoreactivity. 13 All experiments conformed to the Guidelines for Animal Experimentation at Hamamatsu University School of Medicine on the ethical use of animals.…”