Radioimmunoassay for orexin A

Radioimmunoassay for orexin A

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 gu...

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Bibliographic Details
Published in:Life sciences (1973) Vol. 66; no. 10; pp. 897 - 904
Main Authors: Mitsuma, Terunori, Hirooka, Yoshifumi, Kayama, Masato, Mori, Yuichi, Yokoi, Yasutada, Rhue, Nebi, Ping, Jing, Izumi, Masayuki, Ikai, Rumi, Adachi, Koshin, Nogimori, Tsuyoshi
Format: Journal Article
Language:English
Published: Netherlands Elsevier Inc 2000
Subjects:
Animals
Antibody Specificity
Carrier Proteins - analysis
Carrier Proteins - immunology
Carrier Proteins - isolation & purification
Chromatography, Ion Exchange
Immune Sera
Intracellular Signaling Peptides and Proteins
Male
Neuropeptides - analysis
Neuropeptides - immunology
Neuropeptides - isolation & purification
orexin A
Orexins
Rabbits
Radioimmunoassay
rat
Rats
Rats, Wistar
Reproducibility of Results
rat
orexin A
radioimmunoassay
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Summary: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.
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ISSN:0024-3205
1879-0631
DOI:10.1016/S0024-3205(99)00673-6