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|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%|
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.
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.
|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|
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.
|Sandwich ELISA||Use at an assay dependent concentration.|
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.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"