Detection of elastin by immunoelectronmicroscopy. A comparison of different procedures.
Elastin components have been identified in chick aorta by different immunoelectronmicroscopic procedures (peroxidase-antiperoxidase, immunoferritin and immunogold) using affinity purified antibodies to chick tropoelastin. The PAP method used in a preembedding procedure stained the outer portion of amorphous elastin and the microfibrils very intensively. The surface of the cells was also slightly stained. On the contrary immunogold labelling on Epon or Lowicryl embedded sections produced a strong decoration only of amorphous elastin, while microfibrils remained almost completely unlabelled. The result is not due to loss of antigenicity of microfibrils during embedding, since similar data were obtained with immunoferritin in a preembedding procedure. Experiments performed under different stringency conditions showed that the products of the peroxidase reaction diffuse and redistribute in the tissue, indicating that the positive staining of microfibrils and cell surface is an artifact. The value of different immunological reagents and procedures in studying the fine mapping of elastin components is discussed.
Daga-Gordini
D
D
Institute of Histology and Embryology, University of Padova, Italy.
Bressan
G M
GM
Castellani
I
I
Volpin
D
D
D000818:Animals; D001011:Aorta / Q000032:analysis; D002645:Chickens; D004549:Elastin / Q000032:analysis*; D007120:Immunochemistry; D008854:Microscopy, Electron
D004549:Elastin