Human Osteocalcin DuoSet ELISA Summary
* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.
Product Features
- Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
- Development protocols are provided to guide further assay optimization
- Assay can be customized to your specific needs
- Economical alternative to complete kits
Kit Content
- Capture Antibody
- Detection Antibody
- Recombinant Standard
- Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)
Other Reagents Required
DuoSet Ancillary Reagent Kit 3 (5 plates): (Catalog # DY009) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 3.
The components listed above may be purchased separately:
PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered
Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4
Reagent Diluent: (Catalog # DY004), or 5% Tween 20 in PBS, 7.2-7.4, 0.2 μm filtered
Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)
Stop Solution: 2 N H2SO4 (Catalog # DY994)
Microplates: R&D Systems (Catalog # DY990)
Plate Sealers: ELISA Plate Sealers (Catalog # DY992)
Scientific Data
Product Datasheets
Preparation and Storage
Background: Osteocalcin
Osteocalcin, also named Bone gamma-carboxyglutamic Acid Protein, is a secreted protein whose expression is restricted to cells of the osteoblast lineage. It has been frequently used as a marker for osteoblast lineage cells.
Assay Procedure
GENERAL ELISA PROTOCOL
Plate Preparation
- Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
- Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
- Block plates by adding 300 μL of Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
- Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.
Assay Procedure
- Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Repeat the aspiration/wash as in step 2.
- Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
- Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.
Citations for Human Osteocalcin DuoSet ELISA
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
11
Citations: Showing 1 - 10
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Bone Regenerative Effect of Injectable Hypoxia Preconditioned Serum-Fibrin (HPS-F) in an Ex Vivo Bone Defect Model
Authors: Jiang, J;Röper, L;Fuchs, F;Hanschen, M;Failer, S;Alageel, S;Cong, X;Dornseifer, U;Schilling, AF;Machens, HG;Moog, P;
International journal of molecular sciences
Species: Human
Sample Types: Serum
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Surgical Site-Released Tissue Is Potent to Generate Bone onto TCP and PCL-TCP Scaffolds In Vitro
Authors: Rehage, E;Sowislok, A;Busch, A;Papaeleftheriou, E;Jansen, M;Jäger, M;
International journal of molecular sciences
Species: Human
Sample Types: Cell Culture Supernates
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Influence of diabetes mellitus on the pathological profile of aortic stenosis: a sex-based approach
Authors: Martín-Núñez, E;Goñi-Olóriz, M;Matilla, L;Garaikoetxea, M;Mourino-Alvarez, L;Navarro, A;Fernández-Celis, A;Tamayo, I;Gainza, A;Álvarez, V;Sádaba, R;Barderas, MG;Jover, E;López-Andrés, N;
Cardiovascular diabetology
Species: Human
Sample Types: Cell Culture Supernates, Tissue Homogenates
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Calcination and ion substitution improve physicochemical and biological properties of nanohydroxyapatite for bone tissue engineering applications
Authors: Kurzyk, A;Szwed-Georgiou, A;Pagacz, J;Antosik, A;Tymowicz-Grzyb, P;Gerle, A;Szterner, P;W?odarczyk, M;P?oci?ski, P;Urbaniak, MM;Rudnicka, K;Biernat, M;
Scientific reports
Species: Human
Sample Types: Cell Culture Supernates
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Osteoblast-Derived Matrix Vesicles Exhibit Exosomal Traits and a Unique Subset of microRNA: Their Caveolae-Dependent Endocytosis Results in Reduced Osteogenic Differentiation
Authors: Skelton, AM;Cohen, DJ;Boyan, BD;Schwartz, Z;
International journal of molecular sciences
Species: Human
Sample Types: Cell Culture Supernatants
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Methanolic extract of O.umbellata L. exhibits anti-osteoporotic effect via promoting osteoblast proliferation in MG-63�cells and inhibiting osteoclastogenesis in RANKL-stimulated RAW 264.7�cells
Authors: Paramasivam, S;Perumal, SS;
Journal of ethnopharmacology
Species: Human
Sample Types: Cell Culture Supernates
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Association of bone-related�biomarkers with femoral neck bone strength
Authors: N Xia, Y Cai, W Wang, C Bao, Y Li, Q Xie, W Xu, D Liu
Bmc Musculoskeletal Disorders, 2022-05-21;23(1):482.
Species: Human
Sample Types: Serum
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The Brown Algae Ishige sinicola Extract Ameliorates Ovariectomy-Induced Bone Loss in Rats and Suppresses Osteoclastogenesis through Downregulation of NFATc1/c-Fos
Authors: M Kim, M Park
Nutrients, 2022-04-19;14(9):.
Species: Rat
Sample Types: Serum
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Parathyroid Hormone Induces Human Valvular Endothelial Cells Dysfunction That Impacts the Osteogenic Phenotype of Valvular Interstitial Cells
Authors: M Vadana, S Cecoltan, L Ciortan, RD Macarie, AC Mihaila, MM Tucureanu, AM Gan, M Simionescu, I Manduteanu, I Droc, E Butoi
International Journal of Molecular Sciences, 2022-03-29;23(7):.
Species: Human
Sample Types: Cell Culture Supernates
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Osteogenic Differentiation of Human Mesenchymal Stem Cells Modulated by Surface Manganese Chemistry in SLA Titanium Implants
Authors: JW Park, Y Tsutsumi, EK Park
BioMed Research International, 2022-01-13;2022(0):5339090.
Species: Human
Sample Types: Cell Culture Supernates
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Uniaxial Static Strain Promotes Osteoblast Proliferation and Bone Matrix Formation in Distraction Osteogenesis In Vitro
Authors: Z Li, J Zheng, D Wan, X Yang
Biomed Res Int, 2020-08-12;2020(0):3906426.
Species: Rat
Sample Types: Cell Culture Supernates
FAQs
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Are either of R&D Systems Human Osteocalcin ELISA assays, catalog #s DSTCN0 or DY1419, specific for un-carboxylated or gamma-carboxylated form of Osteocalcin?
The Human Osteocaclin Quantikine ELISA (catalog # DSTCN0) assay recognizes natural and recombinant human Osteocalcin, and the assay recognizes Des-γ-carboxylated-Osteocalcin. The Human Osteocalcin DuoSet ELISA (catalog # DY419) also recognizes natural and recombinant human Osteocalcin, however, we have not determined if the DuoSet also recognizes Des-γ-carboxylated-Osteocalcin.
Reviews for Human Osteocalcin DuoSet ELISA
Average Rating: 3.5 (Based on 2 Reviews)
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We used this kit for the quantification of Osteocalcin in human serum and plasma. Worksok and well-described protocol.
