The Ceruloplasmin Activity research-use-only kit is a colorimetric activity assay designed for the quantification and detection of ceruloplasmin activity in serum and urine samples.

This complete, ready-to-use kit includes clear 96 well plate(s), ceruloplasmin standard (200 Units/mL), ceruloplasmin substrate, and other components to perform the assay. A 96-well microplate reader capable of reading optical density at 560 nm is required for use of this kit.

Performance characteristics
• Assay type: colorimetric activity kit
• Sample types: serum and urine samples
• Sensitivity: 3.26 mU/mL
• Standard curve range: 31.2 mU/mL–1,000 mU/mL
• Reactivity: species independent

Background
Ceruloplasmin (Cp) is a multicopper oxidase enzyme involved in the safe handling of oxygen in some metabolic pathways of vertebrates. Discovered in 1948, a blue protein from the a2-globulin fraction of human serum possessing oxidase activity towards aromatic diamines and catechol was purified by Holmberg and Laurell. It was denoted ceruloplasmin, literally meaning 'a blue substance from plasma'. Specialized copper sites have been recruited during evolution to provide long-range electron transfer reactivity and oxygen binding and activation in proteins destined to cope with oxygen reactivity in different organisms.

Ceruloplasmin belongs to the family of multicopper oxidases which are among the few enzymes able to bind molecular oxygen to perform its complete reduction to water. Ceruloplasmin contains 95% of the copper in serum. Cp found in serum is expressed in the liver, but it is also expressed in the brain, lung, spleen, and testis. This assay has been validated for serum and urine samples. Ceruloplasmins are ancient enzymes that should behave in a similar manner to the colorimetric substrate. This assay should to measure Cp activity from a wide range of sources.

Assay principle
The Ceruloplasmin Activity kit is designed to quantitatively measure Ceruloplasmin activity in diluted serum and urine samples. A human ceruloplasmin standard is provided to generate a standard curve for the assay and all samples should be read off of the standard curve. Samples are diluted in the provided Assay Buffer and added to the wells of a half area clear plate. The reconstituted ceruloplasmin substrate is added and the plate is incubated at 30°C for 60 minutes. The ceruloplasmin in the standards and samples reacts with the substrate to produce a colored product. The optical density is read at 560 nm. Increasing levels of ceruloplasmin in the samples causes an increase in the fuchsia (pink-purple) product.

Ceruloplasmin unit definition
Ceruloplasmin activity is based upon the published determination by G. Curzon and L. Vallet, Biochem. J., 1960, 74:279-287. Due to our optimized reagents and incubation times the optical density generated by 1 U/mL of ceruloplasmin is substantially higher than 0.1 OD.

Related links
Learn more about ELISA kits
Learn more about other immunoassays