| Calcitonin (CT) is a 32 amino acid peptide hormone secreted by the parafollicular C-cells of the thyroid gland under serum calcium control. After acute administration, this peptide acts as a potent hypocalcemic and hypophosphatemic hormone by increasing renal calcium clearance and reducing bone resorption. However, its precise physiological role in bone metabolism is not yet fully understood. Various forms of CT may be detected in blood samples, including CT monomer, oxidized monomer, a dimer, and possibly a higher molecular weight precusor of CT. The concentrations of these peptides vary with clinical status, renal function and the tissue origin of CT (normal or ectopic production). Medullary thyroid carcinoma (MTC) is a malignant tumor, developed from the C-cells, secreting calcitonin in large excess. This disease occurs either as a sporadic (80%) or a familial (20%) form, which is transmitted as an autosomal dominant gene or as a component of multiple endocrine neoplasia. |
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| Catalog # | C0115-11F |
| CT may also be secreted in excess by a variety of other tumors such as the oat cell carcinoma of the lung (ectopic production). Moderate hypercalcitoninemia is also observed during pregnancy, in pernicious anemia, renal failure and during the neonatal period. The monomeric form of CT is detected in this assay. |
| Clinical research has documented use of the calcitonin assay in studies of | (a) medullary thyroid carcinoma (MTC), (b) assessing the success of surgery and recurrence of MTC, (c) study of preclinical cases of the familial forms of MTC (MEN II or Sipple syndrome) using the stimulation test (calcium or pentagastrin, and (d) study of the pathophysiology of the calcium-phosphate and bone metabolism. |
| Principle | C0115-11F is a solid phase Enzyme Amplified Sensitivity Immunoassay (EASIA) performed on a microtiter plate for the quantitative measurement of human Calcitonin in human serum and plasma. The assay uses monoclonal antibodies (Mabs) directed against distinct epitopes of human calcitonin. Standards and samples react with the capture monoclonal antibody coated onto the microtiter well and with a monoclonal antibody labeled with horseradish peroxidase (HRP). After an incubation period allowing the formation of a sandwich: coated Mab–human calcitonin – Mab (HRP), the microtiter plate is washed to remove unbound enzyme labeled antibody. Bound enzyme-labeled antibody is measured through a chromogenic reaction. Chromogenic Solution (TMB) is added and incubated. The reaction is stopped with the addition of Stop Solution and the microtiter plate is then read at the appropriate wavelength. The amount of substrate turnover is determined colorimetrically by measuring the absorbance which is proportional to the human calcitonin concentration. A standard curve is plotted and human calcitonin concentrations in samples are determined by interpolation from the standard curve. |
| Specificaions | Sensitivity: 0.5 pg/mL |
| Range: 5 – 950 pg/mL (may vary slightly between lots) |
| Incubation Time: Overnight (18+1 hours) at 4ºC plus 30 minutes at room temperature. |
| Complete: 1 pre-coated strip-well plate. |
| HRP conjugated detection antibody. |
| Dilution buffers, wash solution, TMB and stop solution included. |
| 6 pre-diluted st andards included. |
| 2 plasma controls included. |
| Accurate: Inter-assay CV: 6.3% |
| Intra-assay CV: 3.5% |
| Calibrated to MRC 89/620 reference st andard. |
| Versatile: Serum, EDTA plasma. |
| 50ul sample volume required. |
| Kit Components | C0115-11F1: Standard 1, 0pg/ml. 1x1vial |
| C0115-11F2: Standard 2, 11pg/ml. 1x1vial |
| C0115-11F3: Standard 3, 33pg/ml. 1x1vial |
| C0115-11F4: Standard 4, 135pg/ml. 1x1vial |
| C0115-11F5: Standard 5, 417pg/ml. 1x1vial |
| C0115-11F6: Standard 6, 1146pg/ml. 1x1vial |
| C0115-11F7: Microtiter Plate, 1x96 wells. Coated with monoclonal antibody against human CT |
| C0115-11F8: CT Mab (HRP), 1x1vial. Reconstitute with 125ul Conjugate Buffer. |
| C0115-11F9: Conjugate Buffer, 1x11ml. Supplied as a liquid with preservatives. |
| C0115-11F10: Low Control, 124±39pg/ml, 1x1vial. Reconstitute with 0.5ml distilled water. |
| C0115-11F11: High Control, 490±108pg/ml, 1x1vial. Reconstitute with 0.5ml distilled water. |
| C0115-11F12: CT Free Human Serum, 1x1vial. Reconstitute with 7ml Free Serum Buffer. To be used for sample dilution. |
| C0115-11F113: Free Serum Buffer, 1x8ml. |
| C0115-11F14: Wash Solution, 1x10ml. dilute the vial content in 2000ml distilled water. |
| C0115-11F15. Chromogenic Solution (TMB), 1x25ml. |
| C0115-11F16: Stop Solution, 1x25ml. |
| Note: |
| 1. Standards 1-6 are formulated in CT-free human serum with preservatives and protease inhibitors. Reconstitute with 0.5ml distilled water. |
| 2. 1pg of reference CT preparation is equivalent to 0.19uIU MRC 89/620. |
| 3. Unused strips must be stored at 4°C in the closed bag containing the dessicant. |
| Storage and Stability | Store all kit components at 4°C. Kit stability is 6 months. Store reconstituted Standards, Controls and CT Free Human Serum at -20°C (only 1 free-thaw cycle is recommended). |
| Important Note | Read entire protocol before use. |
| This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological. |
