Osteocalcin ELISA Assay Kit

$450.00

The Eagle Biosciences Human Osteocalcin ELISA Assay Kit (enzyme-linked immunoassay kit) is intended for the quantitative determination of both human osteocalcin (1-49) and osteocalcin (1-43) (also referred as N-terminal & mid-regional osteocalcin) levels in serum or plasma samples. The Eagle Biosciences Human Osteocalcin ELISA Assay Kit is for research use only and not to be used in diagnostic procedures.

SKU: OST31-K01 Categories: , ,

Osteocalcin ELISA Assay Kit

For Research Use Only

Size: 1×96 wells
Sensitivity: 0.31 ng/mL
Dynamic Range: 4.0 – 64 ng/mL
Incubation Time: 1.5 hours
Sample Type: Serum, Plasma
Sample Size: 25 µL
Controls Included

Product Developed and Manufactured in USA

Additional Information

Assay Background

Osteocalcin [also as bone Gla protein (BGP)] is a major noncollagenous protein found in bone and dentin. The synthesis of osteocalcin involves vitamin K and vitamin D3. Freshly synthesized osteocalcin is partly released into the blood stream and partly incorporated into the bone matrix. Both osteocalcin (1-49) and its fragments including osteocalcin (1-43) are released into the blood stream. Serum osteocalcin (1-43) also generated by catabolic breakdown of osteocalcin (1-49) in the circulation, liver, kidney, as well as by in vitro degradation during storage of samples, because a labile six-amino acid C-terminal sequence that, in vitro at room temperature, is easily cleaved off. There are several studies that have confirmed the measurement of the much more stable N-terminal and mid-regional osteocalcin [osteocalcin (1-43/49)] as being clinically useful, which may contribute to a more accurate assessment of the bone turnover rate.

As osteocalcin is manufactured by osteoblasts, it is often used as a biochemical marker, or biomarker, for the bone formation process. It has been routinely observed that higher serum-osteocalcin levels are relatively well correlated with increases in bone mineral density (BMD) during treatment with anabolic bone formation drugs for osteoporosis, such as Forteo. In many studies, Osteocalcin is used as a preliminary biomarker on the effectiveness of a given drug on bone formation.

Assay Procedure

The Eagle Biosciences Human Osteocalcin ELISA Assay Kit is designed, developed and produced for the quantitative measurement of human osteocalcin (1-49) and (1-43) in serum or plasma sample. The assay utilizes the two-site “sandwich” technique with two selected antibodies that bind to different epitopes of human osteocalcin.

Assay standards, controls and samples are added directly to wells of a microtiter plate that is coated with streptavidin. Subsequently, a mixture of biotinylated human osteocalcin N-terminal region specific polyclonal antibody and a peroxidase labeled human osteocalcin 20 – 43 region specific monoclonal antibody is added to each well. After the first incubation period, a “sandwich” of “biotinylated antibody – human osteocalcin – HRP-monoclonal antibody” is formed and this immunocomplex is also captured to the wall of microtiter plate via a streptavidin-biotin affinity binding. The unbound monoclonal antibodies and buffer matrix are removed in the subsequent washing step. A substrate solution in a timed reaction is then measured in a spectrophotometric microplate reader. The enzymatic activity of the immunocomplex bound to the wall of each microtiter well is directly proportional to the amount of human osteocalcin in a test sample. A standard curve is generated by plotting the absorbance versus the respective human osteocalcin concentration for each standard on point-to-point or 4 parameter curve fit. The concentration of human osteocalcin in test samples is determined directly from this standard curve.

  • Place a sufficient number of streptavidin coated microwell strips in a holder to run human Osteocalcin standards, controls and unknown samples in duplicate.

Test Configuration

ROW STRIP 1 STRIP 2 STRIP 3
A STD 1 STD 5 SAMPLE 1
B STD 1 STD 5 SAMPLE 1
C STD 2 STD 6 SAMPLE 2
D STD 2 STD 6 SAMPLE 2
E STD 3 C 1 SAMPLE 3
F STD 3 C 1 SAMPLE 3
G STD 4 C 2
H STD 4 C 2

Typical Standard Curve

osteocalcin-elisa-assay-kit-standard-curve

Manual

Product Manual