Document Detail


Homologous amino-terminal radioimmunoassay for rat parathyroid hormone.
MedLine Citation:
PMID:  1872388     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Existing radioimmunoassays for parathyroid hormone (PTH) in rat plasma are based on cross-reactivity of rat PTH (rPTH) with heterologous antisera. We used the synthetic NH2-terminal fragment of rPTH [rPTH-(1-34)] to develop a homologous radioimmunoassay for circulating PTH. An antiserum to rPTH-(1-34) was raised in a goat (G-813), and the same peptide was used as radioligand (125I) and standard. Purification of the label by high-performance liquid chromatography (HPLC) increased specific binding greater than twofold and sensitivity by 50-100%. With a final antiserum dilution of 1:70,000, maximum specific binding of 30-33%, nonspecific binding of 1-5%, and 50-microliters sample additions, the assay detection limit was 1.8-2.5 pmol/l. A midregional fragment of human PTH did not displace 125I-labeled rPTH-(1-34). HPLC of extracts of rat parathyroid glands and hyperparathyroid plasma showed only a single peak of immunoreactivity that eluted 2 min after rPTH-(1-34). Dose dilution curves for rat parathyroid gland extracts, rPTH-(1-34) added to rat plasma, and endogenous rat plasma PTH all paralleled the standard curve. Immunoreactive PTH (irPTH) was detectable in greater than 90% of fasting normal rat plasma and changed appropriately in response to hyper- and hypocalcemia induced by low-calcium and vitamin D-deficient diets, injections of calcium and EDTA, and after thyroparathyroidectomy. The normal range for rat plasma irPTH was less than 2.0-12 pmol/l, in general agreement with bioassay results of others. Thus rPTH-(1-34) is an excellent immunogen for raising antisera to rPTH, and assays incorporating it may be of great value in studying rat parathyroid physiology.
Authors:
M S Calvo; C M Gundberg; H Heath; J Fox
Related Documents :
8867318 - Methazid-induced vitamin b2 deficiency in rats and its prevention.
3981268 - Urinary lumichrome-level catabolites of riboflavin are due to microbial and photochemic...
3146718 - Enhanced gastrointestinal absorption of aluminium in uraemia: time course and effect of...
1149728 - Effect of excess and deficiency of vitamin a on the utilization of ffa by liver and ske...
6121928 - Effects of in vitro potassium on ammoniagenesis in rat and canine kidney tissue.
9851668 - Effect of adrenalectomy and glucocorticoid therapy on lipid metabolism of lactating rats.
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  The American journal of physiology     Volume:  261     ISSN:  0002-9513     ISO Abbreviation:  Am. J. Physiol.     Publication Date:  1991 Aug 
Date Detail:
Created Date:  1991-09-19     Completed Date:  1991-09-19     Revised Date:  2007-11-15    
Medline Journal Info:
Nlm Unique ID:  0370511     Medline TA:  Am J Physiol     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  E262-8     Citation Subset:  IM    
Affiliation:
Division of Endocrinology and Metabolism, Mayo Clinic, Rochester, Minnesota 55905.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
Animals
Calcium / administration & dosage,  blood
Diet
Evaluation Studies as Topic
Goats
Male
Parathyroid Hormone / blood*,  chemistry
Radioimmunoassay / methods*
Rats / blood*
Rats, Inbred Strains
Species Specificity
Grant Support
ID/Acronym/Agency:
AR-32526/AR/NIAMS NIH HHS
Chemical
Reg. No./Substance:
0/Parathyroid Hormone; 7440-70-2/Calcium

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


Previous Document:  Postnatal changes in lambs of two pathways for thyroxine 5'-monodeiodination in brown adipose tissue...
Next Document:  Cut set analysis of compartmental models with applications to experiment design.