| Project/Area Number |
60570005
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
General anatomy (including Histology/Embryology)
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| Research Institution | Shiga University of Medical Science |
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Principal Investigator |
OCHI Junzo Professor, Department of Anatomy, Shiga University of Medical Science, 医学部, 教授 (10073058)
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| Co-Investigator(Kenkyū-buntansha) |
SAITOH Yasuharu Assistant, Department of Anatomy, Shiga University of Medical Science, 医学部, 助手 (40178512)
KIMURA Hiroshi Assistant Professor, Dept. Anatomy, Shiga University of Med. Sci., 医学部, 助教授 (40079736)
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| Project Period (FY) |
1985 – 1986
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| Project Status |
Completed (Fiscal Year 1986)
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| Budget Amount *help |
¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1986: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1985: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Keywords | Anti-reserpine serum / Immunochemical analysis / 酵素免疫測定法 |
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| Research Abstract |
Since the fundamental element of this project is to produce specific and high titer antiserum against reserpine, the first year supported by the Grant-in-Aid has been occupied by the study for synthesis of suitable immunogen. A detailed chemical analysis indicated that the best carrier protein was bovine serum albumin (BSA) which can be conjugated with haptenic reserpine via glutaraldehyde as a bridge reagent. The optimal reaction for yeilding the conjugate was achieved under the condition as follows; 4mg reserpine, 10 mg BSA, 100 ul of 25% glutaraldehyde and 0.1 M acetate buffer were mixed overnight at pH 4.8. When the conjugate was emulsified with Freund' adjuvant and used for immunizing rabbits, titers of anti-reserpine antibody were gradually detectable,by a newly developed enzyme immunoassay, from fifth injection and reached the maximum at 12th injection. The best serum in both high titer and specificity showed ability to detect less than 3 pmoles of reserpine. Furthermore, absorption tests revealed that the antiserum recognize reserpine specifically not only of BSA-bound form but also of free form. The purpose of the present project was thus almost accomplished by such excellent antiserum. Further study will be promizing for better understanding the role of reserpine on, in particular, central aminergic systems.
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