Ceruloplasmin (aka Ferroxidase I) is a 132kDa 1,046 amino acid glycoprotein which carries 95% of serum copper by binding 6 cupric ions per molecule. Levels are decreased in Wilson’s Disease (hepatolenticular degeneration) and heritable aceruloplasminemia leading to iron accumulation in the liver or brain from impaired iron homeostasis. The sensitive quantitative measurement of human ceruloplasmin in plasma, serum, urine, milk, saliva and cell culture samples is easily performed with this 96 well strip format ELISA kit. The concentration of ceruloplasmin in normal human plasma is 0.3 mg/ml. The assay measures human ceruloplasmin in the 1-1,000 ng/ml range. Samples giving human ceruloplasmin levels above 1000 ng/ml should be diluted in blocking buffer before use. For best results, dilute plasma and serum samples 1:10,000 to 1:50,000 and milk samples 1:10 to 1:50. Saliva and urine samples should be applied directly to the plate for best results. Human ceruloplasmin will bind to the capture antibody coated on the microtiter plate. After appropriate washing steps, biotin labeled anti-human ceruloplasmin primary antibody binds to the captured protein. Excess primary antibody is washed away and bound antibody is reacted with streptavidin conjugated to HRP. Following an additional washing step, TMB substrate is used for color development at 450nm. Color development is proportional to the concentration of ceruloplasmin in the samples. A standard calibration curve is prepared using dilutions of purified ceruloplasmin and is measured along with the test samples. All reagents and standards are provided in these ELISA kits.