Human Osteocalcin Quantikine ELISA Kit

Catalog # Availability Size / Price Qty
DSTCN0
Control Products Available
Human Osteocalcin ELISA Cell Culture Supernate Standard Curve
3 Images
Product Details
Procedure
Citations (9)
FAQs
Supplemental Products
Reviews (1)

Human Osteocalcin Quantikine ELISA Kit Summary

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.

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
Sample 1 2 3 1 2 3
n 4 4 4 4 4 4
Mean 8.15 17.2 33.3 9.06 19.6 40
Standard Deviation 0.268 0.395 0.544 0.306 0.379 0.653
CV% 3.3 2.3 1.6 3.4 1.9 1.6

EDTA Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 4 4 4 4 4 4
Mean 9.06 19.6 40 7.91 16.1 33.6
Standard Deviation 0.306 0.379 0.653 0.766 0.991 1.66
CV% 3.4 1.9 1.6 9.7 6.2 4.9

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 ELISA Linearity

Data Examples

Human Osteocalcin ELISA Cell Culture Supernate Standard Curve

Human Osteocalcin ELISA EDTA Plasma Standard Curve

Product Datasheets

Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
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
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

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 for Human Osteocalcin Quantikine ELISA Kit

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.

9 Citations: Showing 1 - 9
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  1. Transcriptome Analysis of Mesenchymal Stem Cells from Multiple Myeloma Patients Reveals Downregulation of Genes Involved in Cell Cycle Progression, Immune Response, and Bone Metabolism
    Authors: RC Fernando, DR Mazzotti, H Azevedo, AF Sandes, EG Rizzatti, MB de Oliveir, VLF Alves, AIP Eugênio, F de Carvalh, MA Dalboni, DC Martins, GWB Colleoni
    Sci Rep, 2019;9(1):1056.
    Species: Human
    Sample Types: Cell Culture Supernates
  2. 3D Biomimetic Magnetic Structures for Static Magnetic Field Stimulation of Osteogenesis
    Authors: IA Paun, RC Popescu, BS Calin, CC Mustaciosu, M Dinescu, CR Luculescu
    Int J Mol Sci, 2018;19(2):.
    Species: Human
    Sample Types: Cell Culture Supernates
  3. Macroporous alginate foams crosslinked with strontium for bone tissue engineering
    Authors: O Catanzano, A Soriente, A La Gatta, M Cammarota, G Ricci, I Fasolino, C Schiraldi, L Ambrosio, M Malinconic, P Laurienzo, MG Raucci, G Gomez d'Ay
    Carbohydr Polym, 2018;202(0):72-83.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. Weighted Vest Use during Dietary Weight Loss on Bone Health in Older Adults with Obesity
    Authors: JL Kelleher, DP Beavers, RM Henderson, D Yow, C Crotts, J Kiel, BJ Nicklas, KM Beavers
    J Osteoporos Phys Act, 2017;5(4):.
    Species: Human
    Sample Types: Serum
  5. 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 Types: Serum
  6. 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 Types: Serum
  7. Relation between the stability of dental implants and two biological markers during the healing period: a prospective clinical study
    Authors: Choknapa Tirachaimo
    Int J Implant Dent, 2016;2(1):27.
    Species: Human
    Sample Types: Crevicular Fluid
  8. 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 Types: Cell Culture Supernates
  9. Acute myeloid leukemia transforms the bone marrow niche into a leukemia-permissive microenvironment through exosome secretion.
    Authors: Kumar B, Garcia M, Weng L, Jung X, Murakami J, Hu X, McDonald T, Lin A, Kumar A, DiGiusto D, Stein A, Pullarkat V, Hui S, Carlesso N, Kuo Y, Bhatia R, Marcucci G, Chen C
    Leukemia, 0;32(3):575-587.
    Species: Human
    Sample Types: Plasma

FAQs

  1. Why can't heparin plasma samples be used with the Human Osteocalcin Quantikine ELISA Kit, Catalog # DSTCN0? 

    • It was found that heparin tends to accelerate the enzymatic breakdown of osteocalcin and it is therefore not recommended with this kit. 

  2. Why is serum not a validated sample type with the Human Osteocalcin Quantikine ELISA Kit, Catalog # DSTCN0?

    • Serum values have shown to be consistently lower than plasma samples in this kit and therefore are not an ideal sample type. 

  3. 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.

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Reviews for Human Osteocalcin Quantikine ELISA Kit

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Human Osteocalcin Quantikine ELISA Kit
By Anonymous on 05/03/2017
Application: Sample Tested: Human tissue