Radioimmunoassay for hypocretin-2
- PMID: 10808249
Radioimmunoassay for hypocretin-2
Abstract
Objective: To develop radioimmunoassay for hypocretin-2 (Hcrt-2). And search for its presence in certain rat tissues.
Methods: Anti-Hcrt-2 serum has been raised in New Zealand white rabbits immunized with a conjugate of synthetic Hcrt-2 with bovine serum albumin. Radioiodination of Hcrt-2 was performed by chloramine T method, followed by purification of radoiodinated material on Sephadex G-25 column.
Results: The obtained antibody did not cross react with hypocretin-2, hypothalamic hormones, pituitary hormones, neuropeptides or gut hormones. The assay was performed with a double antibody system. Hcrt-2 was extracted from the tissues with acid acetone. The dilution curve of acid acetone extracts of rat hypothalamus in the radioimmunoassay system was parallel to the standard curve. The recovery of tissue Hcrt-2 was about 85 % and the intra-assay and inter-assay variation were 5.6 % and 8.0 %, respectively. Hcrt-2 was found in the hypothalamus, cerebrum, brain stem and testes.
Conclusions: The obtained data suggest that the assay system developed is suitable to measure Hcrt-2 in tissues and that Hcrt-2 is mainly found in the hypothalamus.
Similar articles
-
Radioimmunoassay for aquaporin-9.Endocr Regul. 2004 Dec;38(4):157-60. Endocr Regul. 2004. PMID: 15841795
-
Radioimmunoassay for Aquaporin-2.Endocr Regul. 1998 Sep;32(3):141-144. Endocr Regul. 1998. PMID: 10196511
-
Radioimmunoassay for somatostatin receptor type 2.Endocr Regul. 2001 Mar;35(1):31-4. Endocr Regul. 2001. PMID: 11308994
-
[Chemistry and radioimmunoassy of neuropeptides (author's transl)].No To Shinkei. 1980 May;32(5):489-502. No To Shinkei. 1980. PMID: 6156694 Review. Japanese. No abstract available.
-
An introduction to the immunocytochemical localization of neuropeptides and neurotransmitters.Acta Histochem Suppl. 1981;24:49-77. Acta Histochem Suppl. 1981. PMID: 6165049 Review. No abstract available.
Cited by
-
Increased and decreased muscle tone with orexin (hypocretin) microinjections in the locus coeruleus and pontine inhibitory area.J Neurophysiol. 2001 May;85(5):2008-16. doi: 10.1152/jn.2001.85.5.2008. J Neurophysiol. 2001. PMID: 11353017 Free PMC article.
-
The effect of clomipramine on wake/sleep and orexinergic expression in rats.J Psychopharmacol. 2009 Jul;23(5):559-66. doi: 10.1177/0269881108089606. Epub 2008 Jun 18. J Psychopharmacol. 2009. PMID: 18562438 Free PMC article.