Overview

  • Product name
    Human Osteocalcin ELISA Kit
  • Detection method
    Colorimetric
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    Overall 8 8.54%
    Inter-assay
    Sample n Mean SD CV%
    Overall 3 3.73%
  • Sample type
    Cell culture supernatant, Serum, Plasma
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    0.1 ng/ml
  • Range
    0.156 ng/ml - 10 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 117 114% - 122%
    Heparin Plasma 109 101% - 117%
    EDTA Plasma 108 99% - 119%
    Citrate Plasma 111 107% - 117%
    serum free media 97 90% - 102%

  • Assay time
    1h 30m
  • Assay duration
    One step assay
  • Species reactivity
    Reacts with: Human
  • Product overview

    Osteocalcin in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Osteocalcin protein in human serum, plasma and serum-free cell culture supernatants.


    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.

  • Notes

    Osteocalcin is a small secreted protein 49 amino acids produced by osteoblasts. Secreted OCN has post-translational modification of specific glutamic acid residues via carboxylation to form γ-carboxyglutamtic acid (GLA) residues. This modification is thought to increase the affinity of OCN for free mineral ions in serum, forming a complex, and thus becoming absorbed into the bone extracellular matrix (ECM). OCN also acts as a hormone impacting glucose metabolism and insulin secretion. During bone reabsorption osteoclasts break down the bone matrix releasing bound Osteocalcin into 5 different fragments; N-terminal, Mid, C-terminal, N-terminal-Mid, and Mid-C-terminal into the serum. Circulating levels of intact OCN and OCN fragments have been found to vary significantly between healthy individuals and diseased stats such as Paget’s disease, osteoporosis, fracture, and renal failure.

  • Tested applications
    Suitable for: Sandwich ELISAmore details
  • Platform
    Microplate

Properties

  • Storage instructions
    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Osteocalcin Capture Antibody 1 x 600µl
    10X Osteocalcin Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    ab221825 - Antibody Diluent 5BI 1 x 6ml
    Osteocalcin Human Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS (ab193972) 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Substrate 1 x 12ml
  • Function
    Constitutes 1-2% of the total bone protein. It binds strongly to apatite and calcium.
  • Sequence similarities
    Belongs to the osteocalcin/matrix Gla protein family.
    Contains 1 Gla (gamma-carboxy-glutamate) domain.
  • Post-translational
    modifications
    Gamma-carboxyglutamate residues are formed by vitamin K dependent carboxylation. These residues are essential for the binding of calcium.
  • Cellular localization
    Secreted.
  • Information by UniProt
  • Alternative names
    • BGLAP
    • BGP
    • Bone gamma carboxyglutamate (gla) protein
    • Bone Gla protein
    • Gamma-carboxyglutamic acid-containing protein
    • OC
    • OCN
    • OSTCN_HUMAN
    • Osteocalcin
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab195214 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • Background-subtracted data values (mean +/- SD) are graphed.

  • Interpolated values of Osteocalcin from serum of 10 male donors are plotted.

  • Background-subtracted data values (mean +/- SD, n = 2) are graphed.

  • Background subtracted data from duplicate measurements are plotted. Measureable levels of Osteocalcin were observed in MG-63 (4-day) serum free supernatants, while no Osteocalcin was detected in HeLa supernatants or serum free media.

Protocols

References

This product has been referenced in:
  • Kim HJ  et al. Sequential transfection of RUNX2/SP7 and ATF4 coated onto dexamethasone-loaded nanospheresenhances osteogenesis. Sci Rep 8:1447 (2018). Read more (PubMed: 29362501) »

See 1 Publication for this product

Customer reviews and Q&As

Very quick osteocalcin ELISA

Excellent Excellent 5/5 (Ease of Use)
Abreviews
The kit is extremely easy to use and since there are so few steps (plus pre-coated plates), it takes only 90 minutes.
I used serum samples. Dilution should be at least 1:100 to get OD values within the standard range.
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Verified customer

Submitted Dec 09 2016

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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