Document Detail


Radioimmunoassay for hypocretin-2.
MedLine Citation:
PMID:  10808249     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
T Mitsuma; Y Hirooka; M Kayma; Y Mori; Y Yokoi; M Izumi; N Rhue; J Ping; K Adachi; R Ikai; N Kawai; A Nakayashiki; T Nogimori
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Endocrine regulations     Volume:  34     ISSN:  1210-0668     ISO Abbreviation:  Endocr Regul     Publication Date:  2000 Mar 
Date Detail:
Created Date:  2000-06-13     Completed Date:  2000-06-13     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  9112018     Medline TA:  Endocr Regul     Country:  CZECH REPUBLIC    
Other Details:
Languages:  eng     Pagination:  23-7     Citation Subset:  IM    
Affiliation:
Fourth Department of Internal Medicine Aichi Medical University, Nagakute, Aichi 480-1195, Japan.
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MeSH Terms
Descriptor/Qualifier:
Animals
Antibodies
Antibody Specificity
Brain Chemistry
Brain Stem / chemistry
Hypothalamus / chemistry
Intracellular Signaling Peptides and Proteins
Male
Neuropeptides*
Neurotransmitter Agents / analysis*,  immunology
Organ Specificity
Quality Control
Rabbits
Radioimmunoassay*
Rats
Rats, Wistar
Testis / chemistry
Chemical
Reg. No./Substance:
0/Antibodies; 0/Intracellular Signaling Peptides and Proteins; 0/Neuropeptides; 0/Neurotransmitter Agents; 0/orexins

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