Novel and sensitive noncompetitive enzyme immunoassay for kassinin in rat plasma.
A novel and sensitive noncompetitive enzyme immunoassay for kassinin is described. Kassinin was biotinylated using sulfosuccinimidyl-6-(biotinamido)hexanoate. The biotinylated kassinin was trapped on anti-kassinin IgG-coated polystyrene balls and, after washing to eliminate other biotinylated substances, was eluted with HCl. The biotinylated kassinin eluted was reacted with anti-kassinin Fab'-peroxidase conjugate and trapped onto streptavidin-coated polystyrene balls. Peroxidase activity bound to the polystyrene balls was assayed by fluorimetry. The detection limit of kassinin was 0.13 pg (0.1 fmol)/tube or 0.065 microgram/l of rat plasma, which was 750-fold or 15-fold lower than that for competitive radioimmunoassay.
Tanaka
K
K
Department of Biochemistry, Medical College of Miyazaki, Japan.
Hashida
S
S
Kohno
T
T
Mohri
Z
Z
Murakami
Y
Y
Ishikawa
E
E
D000818:Animals; D001710:Biotin; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G / Q000302:isolation & purification; D015286:Kassinin / Q000097:blood*; D008297:Male; D011863:Radioimmunoassay / Q000379:methods; D051381:Rats; D011919:Rats, Inbred Strains
D007074:Immunoglobulin G; D015286:Kassinin; D001710:Biotin