PUBLICATION
Characterization of osteocalcin (BGP) and matrix Gla protein (MGP) fish specific antibodies: validation for immunodetection studies in lower vertebrates
- Authors
- Simes, D.C., Williamson, M.K., Schaff, B.J., Gavaia, P.J., Ingleton, P.M., Price, P.A., and Cancela, M.L.
- ID
- ZDB-PUB-050113-8
- Date
- 2004
- Source
- Calcified tissue international 74(2): 170-180 (Journal)
- Registered Authors
- Keywords
- none
- MeSH Terms
-
- Amino Acid Sequence
- Animals
- Antibody Specificity/immunology*
- Bone and Bones/immunology*
- Bone and Bones/metabolism
- Calcium-Binding Proteins/immunology*
- Calcium-Binding Proteins/metabolism
- Electrophoresis, Polyacrylamide Gel
- Extracellular Matrix Proteins*
- Fishes/immunology*
- Fishes/metabolism
- Molecular Sequence Data
- Osteocalcin/immunology*
- Osteocalcin/metabolism
- Species Specificity
- Xenopus/immunology*
- Xenopus/metabolism
- PubMed
- 14668966 Full text @ Calcif. Tissue Int.
Citation
Simes, D.C., Williamson, M.K., Schaff, B.J., Gavaia, P.J., Ingleton, P.M., Price, P.A., and Cancela, M.L. (2004) Characterization of osteocalcin (BGP) and matrix Gla protein (MGP) fish specific antibodies: validation for immunodetection studies in lower vertebrates. Calcified tissue international. 74(2):170-180.
Abstract
In fish species the basic mechanisms of bone development and bone remodeling are not fully understood. The classification of bone tissue in teleosts as cellular or acellular and the presence of transitional states between bone and cartilage and the finding of different types of cartilage in teleosts not previously recognized in higher vertebrates emphasizes the need for a study on the accumulation of the Gla-containing proteins MGP and BGP at the cellular level. In the present study, polyclonal antibodies developed against BGP and MGP from A. regius (a local marine teleost fish) and against MGP from G. galeus (a Pacific Ocean shark), were tested by Western blot for their specificity against BGP and MGP from several other species of teleost fish and shark. For this purpose we extracted and purified both proteins from various marine and freshwater teleosts, identified them by N-terminal amino acid sequence analysis and confirmed the presence of gamma-carboxylation in the proteins with the use of a stain specific for Gla residues. Each antibody recognized either BGP or MGP with no cross-reaction between proteins detected. All purified fish BGPs and MGPs tested were shown to be specifically recognized, thus validating the use of these antibodies for further studies.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping