Radioimmunoassay for orexin A.

Abstract:

:A radioimmunoassay for orexin A has been developed. Anti-orexin A antiserum was raised in New Zealand white rabbits immunized with a conjugate of synthetic orexin A with bovine serum albumin. This antibody did not crossreact with orexin B, hypothalamic hormones, pituitary hormones, neuropeptides or gut hormones. Radioiodination of orexin A was performed with the chloramin T method, followed by purification of radioiodinated material on Sephadex G-25 column. Orexin A was extracted from tissues using acid-acetone. The assay was performed with a double antibody system. The dilution curve of acid-acetone-extracts of rat hypothalamus in the radioimmunoassay system was parallel to the standard curve. The recovery of tissue orexin A was about 80%,and the intra-assay and inter-assay variations were 5.2% and 7.8%, respectively. Orexin A was found in the hypothalamus, cerebrum and testis. These data suggest that this assay system is suitable for the measurement of tissue orexin A and that orexin A is found in the central nervous system and testis.

journal_name

Life Sci

journal_title

Life sciences

authors

Mitsuma T,Hirooka Y,Kayama M,Mori Y,Yokoi Y,Rhue N,Ping J,Izumi M,Ikai R,Adachi K,Nogimori T

doi

10.1016/s0024-3205(99)00673-6

subject

Has Abstract

pub_date

2000-01-01 00:00:00

pages

897-904

issue

10

eissn

0024-3205

issn

1879-0631

pii

S0024320599006736

journal_volume

66

pub_type

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