1991 Fiscal Year Final Research Report Summary
Detection of prolactin based on the receptor-binding activity and its application
| Project/Area Number |
02556037
|
|---|
| Research Category |
Grant-in-Aid for Developmental Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
畜産学(含草地学)
|
|---|
| Research Institution | The university of Tokyo |
|---|
Principal Investigator |
SAKAI Senkiti The University of Tokyo, Faculty of Agriculture, Assoc. Prof., 農学部, 助教授 (80114487)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
AOKI Fugaku The University of Tokyo, Faculty of Agriculture, Assist. Prof., 農学部, 助手 (20175160)
KOHMOTO Kaoyu The University of Tokyo, Faculty of Agriculture, Professor, 農学部, 教授 (30011894)
|
|---|
| Project Period (FY) |
1990 – 1991
|
| Keywords | prolactin / radioreceptorassay / radioimmunoasiay / receptor / immobilized-receptor |
|---|
| Research Abstract |
Prolactin sends its hormonal message to target organs through binding with its receptor located on the plasma membrane. Shill et al. reported that the receptor-binding activity of prolactin correlates well with its biological activity. -However, various molecular forms of prolactin have been identified. Among them, for example glycosylatede prolactin is less active than nonglycosylated prolactin. The microsome fraction has been used as a source of the receptor. In the present experiments, we solubilized these membrane proteins with Triton X-100, purified partially using Con-A affinity chromatography, and then immobilized them on a nitrocellulose membrane filter. Using membrane-bound receptor and bovine serum, the serum level of prolactin was determined by radioreceptor assay. The dosedisplacement curve was obtained in the range of 0.4 and 20 ng/ml. Serum could be included in concentrations up to 16% of the assay buffer and, thus the detection limit was about 2.5 ng/ml. Prolactin in the serum was determined by mdioimmunoassay. The ratio of the two assays was nearly 0.99 (r>+O. 99, n=55). The gel filtration profile was identical between the two assays. It was concluded that membranebound receptor can be used for the determination of serum prolactin with satisfactory.
|
