This Human Calcitonin ELISA Kit is based on a competitive immunoassay format. The antigen is captured by antibodies coated on a 96-well plate. A constant concentration of Bt-tracer (biotinylated tracer) and varying concentrations of unlabeled standard or sample peptide compete for binding specifically to the antiserum. Captured Bt-tracer is subsequently bound by SA-HRP (streptavidin-conjugated horseradishperoxidase), which produces a soluble colored or fluorescent product after a substrate is added.
The calcitonin standard is used to make a standard curve in the range 0-25 ng/ml. Standard curves are S-shaped (on a semi-log plot). The measuring range is the range of standard concentrations near the middle or near the IC50 of the standard curve. Unknown sample concentrations are measured by comparing their absorbance with the standard curve.
Sample Type: Formulated for extracted samples
Assay Range: 0.02-25ng/ml
Average IC50: 1ng/ml
Kit Components:
C0115-12A: Microtiter Plate, 1x96 wells
C0115-12B: EIA Buffer Concentrate (20X), 1x50ml
C0115-12C: Antiserum, lyophilized powder.
C0115-12D: Standard, 1x1ug. Reconstitute with EIA buffer.
H-Cys-Gly-Asn-Leu-Ser-Thr-Cys-Met-Leu-Gly-Thr-Tyr-Thr-Gln-Asp-Phe-Asn-Lys-Phe-His-Thr-Phe-Pro-Gln-Thr-Ala-Ile-Gly-Val-Gly-Ala-Pro-NH2, (Disulfide bond)
C0115-12E: Biotinylated Tracer, lyophilized powder
C0115-12F: Streptavidin-HRP Concentrate (200X). 1x100ul
C0115-12G: TMB Substrate Solution, 1x11ml. Mixture of TMB and H2O2
C0115-12H: Stop Solution, 1x15ml (2N HCl)
Storage and Stability:
Store C0115-12C (Antiserum) and C0115-12D (Standard) at -20 degrees C. Store all the other kit components at 4 degrees C. Kit is stable for up to 6 months.
Cross-reactivity:
Peptide Percent (%)
Calcitonin (human) 100
Calcitonin (rat) 5.4
Calcitonin C-terminal adjacent Peptide (rat) 0.001
Calcitonin (eel) 0
Calcitonin (salmon I) 0
Calcitonin (chicken) 0
Amylin (human) 0
Alpha-CGRP (human) 0
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