Assay Principle:

The DetectX® Ceruloplasmin Colorimetric Activity Kit provides a reliable method to quantitatively measure ceruloplasmin activity in serum and urine samples. This colorimetric activity assay, with a run time of 60 minutes, is designed for efficient and precise analysis. Read the complete kit insert thoroughly before conducting the assay. The kit includes a Ceruloplasmin Standard to establish an accurate standard curve.

Protocol Summary:

• Introduce standards and samples into the provided transparent microtiter plate.
• Add Assay Buffer and reconstituted Cp Substrate solution to each well, ensuring proper mixing.
• Incubate the plate at 30°C for 60 minutes. The color-generating reaction occurs between the Ceruloplasmin Substrate and Ceruloplasmin in the samples.
• After incubation, use a plate reader to detect the signal at 560nm. Calculate Ceruloplasmin activity using the intensity and the standard curve.

Background:

Ceruloplasmin (Cp) is a multifaceted acute phase multicopper oxidase enzyme that plays a protective role in immune responses. Cp is intricately linked with reproductive processes. Estrogen significantly influences Cp production, as it alters the subcellular distribution of copper in the liver, leading to increased plasma copper levels and enhanced ceruloplasmin synthesis.

The elevation of Cp serum levels during normal pregnancy in certain species like humans and other mammals indicates its potential protective function against oxidative stress during reproduction. Evidence supports that the increase in Cp levels may safeguard both the mother and developing fetus from oxidative damage, highlighting Cp’s importance in fetal development and maternal health.

Alternatively, in giant pandas and some felid species, urinary Cp activity shows a marked increase in pregnant versus pseudopregnant animals, observable from the early stages of gestation and continuing throughout the luteal phase.

The DetectX® Ceruloplasmin Colorimetric Activity Kit is a valuable tool for researchers and clinicians in the fields of reproductive biology, endocrinology, and immunology, enabling the accurate measurement of ceruloplasmin activity in various biological samples.