Detection of islet cell specificity of monoclonal islet cell surface antibodies by means of double-staining immunofluorescence.
We have generated monoclonal antibodies (mcab) reactive with islet cell surface antigens. 10 different mcab were characterized regarding their islet cell binding specificity by means of a modified double immunofluorescence test. At this assay, the monoclonal islet cell surface antibodies were visualized on rat islet cells by indirect immunofluorescence with fluorescein isothiocyanate-labelled antibodies against mouse immunoglobulin. The alpha and beta cell specificity was determined by indirect immunofluorescence using anti-glucagon or anti-insulin serum and a tetramethyl rhodamine isothiocyanate-labelled second antibody. The target islet cell suspension used contained 61% beta and 23% alpha cells. The monoclonal antibody K28D6 preferentially reacted with alpha cells. The binding of K29aC6 and K56aF3 indicates a high specificity against beta cells. The remaining 7 antibodies were reactive with alpha as well as with beta cells.
Witt
S
S
Central Institute of Diabetes Gerhard Katsch, Karlsburg, GDR.
Ziegler
B
B
Ziegler
M
M
D000818:Animals; D000911:Antibodies, Monoclonal; D000954:Antigens, Surface / Q000032:analysis*; D005455:Fluorescent Antibody Technique; D005456:Fluorescent Dyes; D007515:Islets of Langerhans / Q000166:cytology*; D051381:Rats; D011919:Rats, Inbred Strains
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D005456:Fluorescent Dyes