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Specific radioimmunoassay for ovine bone gla-protein (osteocalcin)

Journal Article:

Abstract

We developed a sensitive and specific radioimmunoassay for ovine bone gla-protein (osteocalcin) using a polyclonal rabbit antibody raised against ovine bone gla-protein. Bone from lambs was extracted in 0.5 mol/l EDTA and desalted on Sephadex G-25. Bone gla-protein was purified by gel filtration chromatography over Sephadex G-100 and ion-exchange chromatography on DEAE-Sephadex A-25. The protein, subjected to monodimensional electrophoresis migrated as a single spot in SDS PAGE with the same apparent molecular weight of 12 kD as bovine bone gla-protein. The amino acid composition of pufified bone gla-protein was in agreement with a previous publication. The competitive RIA uses /sup 125/I-labelled bone gla-protein as a tracer and a complex of a second antibody and polyethylene glycol to separate free and antibody-bound /sup 125/I-labelled bone gla-protein. The intra- and inter-assay variations are less than 6 and 10%, respectively. There is no reactivity of our antisera with dog sera. The cross-reactivity is only partial with calf and human sera and complete with ovine sera. We measured bone gla-protein levels in serum of 96 normal male sheep of different ages. Serum bone gla-protein rapidly and significantly (P<0.001) decreased from 532 +- 169 ..mu..g/l at birth, to 240 +- 43 ..mu..g/l at 45 days,  More>>
Publication Date:
Jan 01, 1988
Product Type:
Journal Article
Reference Number:
NORN-88-000172; EDB-89-004234
Resource Relation:
Journal Name: Acta Endocrinol.; (Denmark); Journal Volume: 119
Subject:
60 APPLIED LIFE SCIENCES; 63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.; PROTEINS; GLUTAMIC ACID; RADIOIMMUNOASSAY; OSTEOPOROSIS; BONE CELLS; GEL PERMEATION CHROMATOGRAPHY; IODINE 125; SHEEP; SPECIFICITY; AMINO ACIDS; ANIMAL CELLS; ANIMALS; BETA DECAY RADIOISOTOPES; CARBOXYLIC ACIDS; CHROMATOGRAPHY; CONNECTIVE TISSUE CELLS; DAYS LIVING RADIOISOTOPES; DISEASES; DOMESTIC ANIMALS; ELECTRON CAPTURE RADIOISOTOPES; IMMUNOASSAY; IMMUNOLOGY; INTERMEDIATE MASS NUCLEI; IODINE ISOTOPES; ISOTOPE APPLICATIONS; ISOTOPES; MAMMALS; NUCLEI; ODD-EVEN NUCLEI; ORGANIC ACIDS; ORGANIC COMPOUNDS; RADIOASSAY; RADIOIMMUNOLOGY; RADIOISOTOPES; RUMINANTS; SEPARATION PROCESSES; SKELETAL DISEASES; SOMATIC CELLS; TRACER TECHNIQUES; VERTEBRATES; 553006* - Agriculture & Food Technology- Other Agricultural Applications- (1987-); 560162 - Radionuclide Effects, Kinetics, & Toxicology- Animals, Plants, Microorganisms, & Cells
OSTI ID:
6700157
Research Organizations:
Unite INSERM 234, Hopital E. Herriot, Lyon (FR)
Country of Origin:
Denmark
Language:
English
Other Identifying Numbers:
Journal ID: CODEN: ACENA
Submitting Site:
DKN
Size:
Pages: 152-160
Announcement Date:
May 01, 1988

Journal Article:

Citation Formats

Pastoureau, P, Merle, B, and Delmas, P D. Specific radioimmunoassay for ovine bone gla-protein (osteocalcin). Denmark: N. p., 1988. Web.
Pastoureau, P, Merle, B, & Delmas, P D. Specific radioimmunoassay for ovine bone gla-protein (osteocalcin). Denmark.
Pastoureau, P, Merle, B, and Delmas, P D. 1988. "Specific radioimmunoassay for ovine bone gla-protein (osteocalcin)." Denmark.
@misc{etde_6700157,
title = {Specific radioimmunoassay for ovine bone gla-protein (osteocalcin)}
author = {Pastoureau, P, Merle, B, and Delmas, P D}
abstractNote = {We developed a sensitive and specific radioimmunoassay for ovine bone gla-protein (osteocalcin) using a polyclonal rabbit antibody raised against ovine bone gla-protein. Bone from lambs was extracted in 0.5 mol/l EDTA and desalted on Sephadex G-25. Bone gla-protein was purified by gel filtration chromatography over Sephadex G-100 and ion-exchange chromatography on DEAE-Sephadex A-25. The protein, subjected to monodimensional electrophoresis migrated as a single spot in SDS PAGE with the same apparent molecular weight of 12 kD as bovine bone gla-protein. The amino acid composition of pufified bone gla-protein was in agreement with a previous publication. The competitive RIA uses /sup 125/I-labelled bone gla-protein as a tracer and a complex of a second antibody and polyethylene glycol to separate free and antibody-bound /sup 125/I-labelled bone gla-protein. The intra- and inter-assay variations are less than 6 and 10%, respectively. There is no reactivity of our antisera with dog sera. The cross-reactivity is only partial with calf and human sera and complete with ovine sera. We measured bone gla-protein levels in serum of 96 normal male sheep of different ages. Serum bone gla-protein rapidly and significantly (P<0.001) decreased from 532 +- 169 ..mu..g/l at birth, to 240 +- 43 ..mu..g/l at 45 days, 152 +- 44 ..mu..g/l at 90 days, and 5.9 +- 0.7 ..mu..g/l at 7 years age. In addition, bone gla-protein levels at birth were higher in normal birth weight than in hypotrophic lambs with low birth weight (535 +- 169 vs 271 +- 156 ..mu..g/l, P<0.0001). Furthermore, lambs raised outside in free conditions tended to have higher serum bone gla-protein levels than lambs raised under shelter (1984 +- 53 vs 137 +- 34 ..mu..g/l), suggesting a role of breeding factors such as diet or relative immobilization on bone gla-protein levels. (Abstract Truncated)}
journal = {Acta Endocrinol.; (Denmark)}
volume = {119}
journal type = {AC}
place = {Denmark}
year = {1988}
month = {Jan}
}