Human Anti-CaSR ELISA Assay Kit (enzyme-linked immunoassay kit) kit is produced for the quantitative determination of human anti-CaSR (calcium sensing receptor) autoantibody levels in serum, plasma, tissue extract or other liquid samples.
Optimal working dilution should be determined by the investigator.
Volume d'échantillon
100 μL
Durée du test
2 h
Protocole
Human Anti-CaSR ELISA Assay Kit designed, developed and produced for the quantitative measurement of human anti-CaSR autoantibody (IgG type) in test samples. The assay utilizes the enzyme linked immunoabsorbent technique with selected immunogenic extracellular CaSR antigen and HRP labeled human IgG specific detection antibody. Assay standards, controls and prediluted patient samples are added to microtiter wells of a microplate which is coated with a highly purified human CaSR extracellular antigen. After the first incubation period, the CaSR antigen on the wall of microtiter well absorbs or captures human anti-CaSR autoantibody in the sample and unbound proteins in each microtiter well are washed away. Then a HRP conjugated polyclonal anti-human IgG antibody is added to each microtiter well and a link of "CaSR antigen - human anti-CaSR autoantibody - HRP conjugated detection antibody" is formed. The unbound detection antibody is removed in the subsequent washing step. HRP conjugated detection antibody bound to the well is then incubated with a substrate solution in a timed reaction and then measured in a spectrophotometric microplate reader. The enzymatic activity of the detection antibody bound to the human anti-CaSR autoantibody on the wall of the microtiter well is directly proportional to the amount of this autoantibody in the sample. A standard curve is generated by plotting the absorbance versus the respective autoantibody concentration for each standard on point-to-point or cubical scales. The concentration of human anti-CaSR autoantibody in test samples is determined directly from this standard curve.