Mid-Tact Human Osteocalcin EIA Kit

About The Assay

Osteocalcin (5800 daltons) is a specific product of the osteoblast.  Only a small amount of osteocalcin is released directly into the circulation, it is primarily deposited into the bone matrix.  Studies have shown that osteocalcin circulates both as the intact (1-49) protein and as N-terminal fragments.  The major N-terminal fragment is the peptide (1-43).

The BTI Mid-Tact Osteocalcin Elisa Kit is highly specific. It measures both the intact human osteocalcin and the major (1-43) fragment.  The assay is a sandwich EIA which employs two monoclonal antibodies.  One antibody (1-19) is immobilized in the wells and the second antibody (30-40) is biotinylated.  The assay is highly sensitive (0.5ng/ml) and requires only a 25 microliter sample.  All the necessary reagents, a 96-well strip plate, and a complete 3½ hour protocol are included with the kit.

References

1.  Garnero P., Grimaux M., Seguin P. and P.D. Delmas. Characterization of Immunoreactive Forms of Human Osteocalcin in vivo and in vitro. J. of Bone & Mineral Res. 9 (2):  255-264, l994.

2.   Calvo M.S., Eyre D.R.,and Gundberg C.M.  Molecular Basis and Clinical Application of Biological Markers of Bone Turnover. Endocrine Reviews 17(4): 333-364, 1996.

Procedural Outline

1.  Add 25ul standard/sample. Add antibody and gently swirl. Incubate 2½ hours at 37^oC.

2.  Aspirate and wash 3 times with wash buffer.

3.  Add Strep-Av-HRP, swirl, incubate 30 mins. at room temperature.

4.  Repeat Step 2. Then add 100ul of substrate, swirl, develop 15 minutes in the dark.
     Add 100ul stop solution, swirl, measure absorbance at 450nm.

BTI Mid-Tact Human Osteocalcin EIA Kit
Catalog No:  BT-480
(sufficient reagents and precoated 96-well strips)
Quantity:  1 kit

Specifications:
Sample Size:  25ul
Assay Time:  3.5 hours
Sensitivity:  0.5 ng/ml
Range:  1.0-50 ng/ml
Intraassay Variation:  7%
Interassay Variation:  10%

FOR RESEARCH USE ONLY.  NOT FOR USE IN HUMANS OR AS AN IN-VITRO DIAGNOSTIC.