TY - JOUR
T1 - Localization of angiotensin converting enzyme (Kininase II). I. Preparation of antibody heme octapeptide conjugates
AU - Yotsumoto, Hideki
AU - Sato, Shoichiro
AU - Shibuya, Masabumi
N1 - Funding Information:
Acknowledgements This work was supportedb y grantsf rom the U.S. Public Health Service( HL 15691,H L 16407, AM-09001-10, and Contract NO1 HR3-3015), from the John A. Hartford Foundation, Inc., the Council for Tobacco Research-U.S.A., Inc., and by an Established Investigatorshipa ward to Dr Una S. Ryan from the American Heart Association and with funds contributed by the Heart Association of Palm BeachC ounty, Florida, and by the VeteransA dministration (Project no. 7963-01).
PY - 1976
Y1 - 1976
N2 - Antibodies to pig lung angiotensin converting enzyme (kininase II) were conjugated to a heme octapeptide (8 microperoxidase, 8 MP) derived from cytochrome c. 8 MP, which has only one reactive amine, was coupled to antibody in a twostep procedure using a bifunctional active ester, bis succinyl succinate. In the first step, 8 MP was reacted with an excess of bis succinyl succinate to yield 8 MP succinyl succinate, a stable compound which can be stored. In a second step the remaining active ester was used for coupling to reactive amines of the antibody. The conjugate consists of 1.6-2.3 8 TP moieties per antibody. Using these procedures, the formation of complex polymers is avoided. Each molecular of conjugate possesses both immunoreactivity and peroxidatic activity. The conjugate has been used to localize angiotensin converting enzyme along the plasma membrane and associated caveolae of pig aortic endothelial cells in culture.
AB - Antibodies to pig lung angiotensin converting enzyme (kininase II) were conjugated to a heme octapeptide (8 microperoxidase, 8 MP) derived from cytochrome c. 8 MP, which has only one reactive amine, was coupled to antibody in a twostep procedure using a bifunctional active ester, bis succinyl succinate. In the first step, 8 MP was reacted with an excess of bis succinyl succinate to yield 8 MP succinyl succinate, a stable compound which can be stored. In a second step the remaining active ester was used for coupling to reactive amines of the antibody. The conjugate consists of 1.6-2.3 8 TP moieties per antibody. Using these procedures, the formation of complex polymers is avoided. Each molecular of conjugate possesses both immunoreactivity and peroxidatic activity. The conjugate has been used to localize angiotensin converting enzyme along the plasma membrane and associated caveolae of pig aortic endothelial cells in culture.
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U2 - 10.1016/0028-2243(76)90082-4
DO - 10.1016/0028-2243(76)90082-4
M3 - Article
C2 - 178067
AN - SCOPUS:84886617409
VL - 8
SP - 111
EP - 124
JO - Scientific Computing and Instrumentation
JF - Scientific Computing and Instrumentation
SN - 1078-8956
IS - 1
ER -