Immunodetection of secretogranin II in animal and human tissues by new monoclonal antibodies.
Secretogranin II (chromogranin C) is an acidic tyrosine-sulfated secretory protein, known to be a marker of neuroendocrine secretory products and of specific neuroendocrine tumours. In order to obtain anti-secretogranin II monoclonal antibodies for cell biology studies and, in particular, for clinical applications, we immunized mice with a secretogranin II-enriched fraction prepared from homogenates of bovine anterior pituitaries. Hybridoma supernatants obtained from the splenocytes of a hyperimmune mouse, screened with an enzyme-linked immunosorbent assay, were analyzed by both immunocytochemistry and two-dimensional immunoblotting. By using this experimental approach, we were able to identify two monoclonal antibodies (8G1 and 5A7) which recognize bovine secretogranin II. Both immunocytochemistry and immunoblotting revealed that one of them, the 5A7 antibody, cross-reacts with the human antigen. The distribution patterns of the immunoreactivity, obtained by immunocytochemistry with the 5A7 antibody in animal and human tissues, partially overlap those, obtained by using a polyclonal antiserum elicited against bovine secretogranin II, previously described. Moreover, the 5A7, but not the polyclonal antibody, reacts with some duodeno-jejunal cells. In conclusion, both the 5A7 and 8G1 antibodies can be useful for cell biology studies. The 5A7 antibody can be used for the detection of secretogranin II in human tissues and should be of help in clinical and pathological practices.
Pelagi
M
M
Istituto Scientifico Ospedale S. Raffaele, Milano, Italy.
Zanini
A
A
Gasparri
A
A
Ermellino
L
L
Giudici
A M
AM
Ferrero
S
S
Siccardi
A G
AG
Buffa
R
R
D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D002415:Cats; D002417:Cattle; D002864:Chromogranins; D003429:Cross Reactions; D004285:Dogs; D006168:Guinea Pigs; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D015151:Immunoblotting; D007150:Immunohistochemistry; D007413:Intestinal Mucosa / Q000378:metabolism; D010903:Pituitary Gland, Anterior / Q000378:metabolism; D011506:Proteins / Q000276:immunology* / Q000378:metabolism*; D051381:Rats; D013045:Species Specificity
D000911:Antibodies, Monoclonal; D002864:Chromogranins; D011506:Proteins