CALPROTECTIN ELISA KIT
INTENDED USE
The Calprotectin ELISA Kit is intended for quantitative determination of Calprotectin (MRP8/14) in stole. It is for in vitro diagnostic use only.
INTRODUCTION
Calprotectin is a calcium-binding protein secreted predominantly by neutrophils and monocytes. Fecal Calprotectin is a marker for neoplasic and inflammatory gastrointestinal diseases.
It is often difficult to distinguish between irritable bowel syndrome and chronic inflammatory bowel disease. This leads in many cases to extensive and unnecessary colonoscopic examinations. The Calprotectin test allows clear differentiation between the two patient groups. Fecal Calprotectin levels correlate significantly with histologic and endoscopic assessment of disease activity in Morbus Crohn's disease and ulcerative colitis as well as with the fecal excretion of indium-111-labelled neutrophilic granulocytes that has been suggested as the “gold standard“ of disease activity in inflammatory bowel disease. However, measuring 111-indium-labeled granulocytes is very costly (patient’s hospitalization, analysis and disposal of isotopic material) and is connected with radioactive exposition of the patients. For this reason, a repeated application to children and pregnant women is not recommended.
Elevated levels of Calprotectin are a much better predictor of relapse than standard inflammatory markers (CRP, ESR HB). Comparing this marker with standard fecal occult blood screening in colorectal cancer demonstrates clearly the diagnostic advantages of the fecal Calprotectin test. The parameter is of a high diagnostic value: if the Calprotectin level in stool is low, the probability is high that no organic intestinal disease exists.
PRINCIPLE OF THE TEST
Calprotectin ELISA Kit
Calprotectin ELISA Kit is intended for quantitative determination of Calprotectin (MRP8/14) in stole. The Calprotectin ELISA Kit assay utilizes the two-site “sandwich” technique with two selected monoclonal antibodies that bind to human Calprotectin.
Standards, controls and diluted patient samples which are assayed for human Calprotectin are added to wells of microplate coated with a high affine monoclonal anti-human Calprotectin antibody. During the first incubation step, Calprotectin in the samples is bound by the immobilized antibody. In a next incubation step, a biotinylated monoclonal anti-human Calprotectin antibody is added to each microtiter well. Then a peroxidase labeled exravidin conjugate is added to each well and the following complex is formed: capture antibody - human Calprotectin – biotinylated detection antibody - Peroxidase conjugate. Tetramethylbenzidine (TMB) is used as a substrate for peroxidase. Finally, an acidic stop solution is added to terminate the reaction. The color changes from blue to yellow. The intensity of the yellow color is directly proportional to the Calprotectin concentration of sample. A dose response curve of the absorbance unit (optical density, OD at 450 nm) vs. concentration is generated, using the values obtained from standard. Calprotectin present in the patient samples, is determined directly from this curve.
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