Osteocalcin antibody | AbD02591

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Human anti Bovine Osteocalcin

Product Type
Monoclonal Antibody
Clone
AbD02591
Isotype
HuCAL Fab bivalent
Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
HCA029 E WB 50 µg
Human anti Bovine osteocalcin antibody, clone AbD02591 recognizes bovine osteocalcin (Bone Gla Protein), a 49 amino acid single chain vitamin K dependent protein (molecular weight 5.8 kDa), made by osteoblasts and a major component of the non-collagenous bone matrix. Post-translational modification by a vitamin K dependent carboxylase produces three g-carboxyglutamic acid residues at positions 17, 21 and 24, giving it a high affinity for calcium. The mature protein contains a single intrachain disulfide bond joining Cys23 to Cys29. The secondary structure is highly calcium dependent and contains 14% a-helix, 20% ß-sheet and 67% random form in the presence of calcium, and 1% a-helix, 20% β-sheet and 79% random form in the ab-sence of calcium (Delmas et al. 1984). Sixty to ninety percent of de novo synthesized osteocalcin is incorporated into the bone matrix where it binds to hydroxy-apatite during matrix mineralization. The remainder of the osteocalcin is released into the circulation where it can be measured as a sensitive marker of bone formation. Serum osteocalcin is elevated in diseases characterized by increased bone turnover such as osteoporosis, hyperparathyroidism and Paget’s disease, and low in conditions associated with low bone turnover such as hypoparathyroidism and growth hormone deficiency ( Lee et al. 1990). Circulating osteocalcin is unstable as it contains a tryptic cleavage site at amino acids 43-45 near the C-terminus. After cleavage a large fragment containing amino acids 1 – 43 is formed, which contains both the N-terminus and the middle portion of the protein. Whereas the levels of intact osteocalcin rapidly decreases, 1-43 osteocalcin remains stable in serum even after repeated freeze thaw cycles or storage at elevated temperatures ( Lee et al. 1990 for details.
Human anti Bovine osteocalcin antibody, clone AbD02591 recognizes intact osteocalcin but not fragments corresponding to amino acids 7-19, 37-49 or 45-49.

Product Details

Target Species
Bovine
Species Cross-Reactivity
Target SpeciesCross Reactivity
Human
N.B. Antibody reactivity and working conditions may vary between species.
Product Form
A bivalent human recombinant Fab selected from the HuCAL® GOLD phage display library. Expressed in E. coli and purified using NiNTA affinity chromatography. This Fab fragment is dimerized via a helix-turn-helix motif. The antibody is tagged with a myc-tag (EQKLISEEDL) and a his-tag (HHHHHH) at the C-terminus of the anti-body heavy chain - Lyophilized.
Reconstitution
Reconstitute with 50 ul distilled water
Care should be taken during reconstitution as the protein may appear as a film at the bottom of the vial. Bio-Rad recommend that the vial is gently mixed after reconstitution.
Preparation
Purified antibody prepared by metal chelate affinity chromatography
Buffer Solution
Phosphate buffered saline
Preservative Stabilisers
None present
Immunogen
Native bovine osteocalcin with the sequence YLDHWLGAPAPYPDPLEPKREVCELNPDCDELADHIGFQEAYRRFYGPV
Approx. Protein Concentrations
Antibody concentration 1.0 mg/ml following reconstitution

Storage Information

Storage
Prior to reconstitution store at +4oC.
After reconstitution store at -20oC.
Storage in frost-free freezers is not recommended. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
Shelf Life
6 months from date of reconstitution

More Information

UniProt
PO2820 Related reagents
Acknowledgements
Sold under license of U.S. Patents 6753136, 7785859 and 8273688 and corresponding patents. This antibody was developed by Bio-Rad, Zeppelinstr. 4, 82178 Puchheim, Germany.
His-tag is a registered trademark of EMD Biosciences.
Licensed Use
For in vitro research purposes only, unless otherwise specified in writing by Bio-Rad.
Regulatory
For research purposes only

Applications of Osteocalcin antibody

This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.
Application Name Verified Min Dilution Max Dilution
ELISA 2 ug/ml
Western Blotting 5 ug/ml
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
Technical Advice
Recommended protocols and further information about HuCAL recombinant antibody technology can be found in the HuCAL Antibodies Technical Manual

Secondary Antibodies Available

Description Product Code Pack Size Applications List Price Quantity
Goat anti Human IgG F(ab')2:HRP 0500-0099 0.5 ml E WB
Mouse anti c-Myc:HRP MCA2200P 0.1 mg C E P WB*
Mouse anti Penta Histidine Tag:HRP MCA5995P 125 µl E WB

Product Specific References

References for Osteocalcin antibody

  1. Delmas PD et al. (1984) Immunochemical studies of conformational alterations in bone gamma-carboxyglutamic acid containing protein.
    Biochemistry. 23: 4720-4725.
  2. Lee, A.J. et al. (2000) Measurement of osteocalcin
    Ann Clin Biochem. 37: 432-446.