• Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    4.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (50 uL), EDTA Plasma (50 uL)
  • Sensitivity
    0.898 ng/mL
  • Assay Range
    2.0 - 64 ng/mL (Cell Culture Supernates, EDTA Plasma)
  • Specificity
    Natural and recombinant human Osteocalcin.
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    No significant interference observed with available related molecules.
Control Available
QC137, Quantikine Immunoassay Control Set 823 for Human Osteocalcin - Please Inquire
Product Summary
The Quantikine Human Osteocalcin Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human Osteocalcin in cell culture supernates and EDTA plasma. It contains synthetic human Osteocalcin and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human Osteocalcin showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring human Osteocalcin.

Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components.
Cell Culture Supernates
Intra-Assay Precision Inter-Assay Precision
Sample123123
n444444
Mean8.1517.233.39.0619.640
Standard Deviation0.2680.3950.5440.3060.3790.653
CV%3.32.31.63.41.91.6

EDTA Plasma
Intra-Assay Precision Inter-Assay Precision
Sample123123
n444444
Mean9.0619.6407.9116.133.6
Standard Deviation0.3060.3790.6530.7660.9911.66
CV%3.41.91.69.76.24.9

Recovery

The recovery of human Osteocalcin spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 105 98-120
EDTA Plasma (n=4) 102 80-116
Linearity
To assess the linearity of the assay, samples spiked with high concentrations of human Osteocalcin were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human Osteocalcin Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
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.
    • Long Name
      Bone gamma-Carboxyglutamate [gla] Protein
    • Entrez Gene IDs
      632 (Human); 12096 (Mouse); 25295 (Rat);
    • Alternate Names
      BGLAP; BGP; bone gamma-carboxyglutamate (gla) protein (osteocalcin); bone gamma-carboxyglutamate (gla) protein; Bone Gla protein; Gamma-carboxyglutamic acid-containing protein; OC; OCN; Osteocalcin;
    Related Research Areas
    Assay Procedure
    Refer to the product for complete assay procedure.

    Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
    1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
    2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

    3. 100 µL Assay Diluent
    4.   Add 100 µL of Assay Diluent to each well.

    5. 50 µL Standard, Control, or Sample
    6.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
    7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

    8. 200 µL Conjugate
    9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
    10.   Aspirate and wash 4 times.

    11. 200 µL Substrate Solution
    12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

    13. 50 µL Stop Solution
    14.   Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
    Citations:

    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.

    3 Citations: Showing 1 - 3
    Filter your results:

    Species
    Sample Type
    1. Undercarboxylated osteocalcin reverts insulin resistance induced by endoplasmic reticulum stress in human umbilical vein endothelial cells
      Authors: Q Guo, H Li, L Xu, S Wu, H Sun, B Zhou
      Sci Rep, 2017;7(1):46.
      Species: Human
      Sample Type: Serum
    2. Identification of suitable reference gene and biomarkers of serum miRNAs for osteoporosis
      Sci Rep, 2016;6(0):36347.
      Species: Primate - Macaca mulatta (Rhesus Macaque)
      Sample Type: Serum
    3. In vitro and in vivo evaluation of zinc-modified ca-si-based ceramic coating for bone implants.
      Authors: Yu J, Li K, Zheng X, He D, Ye X, Wang M
      PLoS ONE, 2013;8(3):e57564.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    ELISA Controls
    Description Application Cat# Citations Images  

    Quantikine Immunoassay Control Set 823 for Human Osteocalcin

    Ctrl QC137
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    Supplemental ELISA Products
    Description Application Cat# Citations Images  

    Quantikine ELISA Wash Buffer 1

    ELISA WA126 6
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    Average Rating: 5 (Based on 1 review)

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    Sample TestedHuman tissue

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