Unconjugated Bilirubin Quantification Kit is designed for easy and direct measurement of unconjugated bilirubin (UCB) in biological specimens. The assay provides specific and accurate quantification of UCB at concentrations as low as 1-5 nM in 50 µL of the sample. This sensitivity is 1,000 times higher than that of commonly used colorimetric Diazo- method. The assay is also free from common for colorimetric method interferences such as pigments, lipids, heme, and hemoglobin.
In biological fluids:
– Quantification of UCB in small-volume samples;
– Quantification of low levels of UCB in the blood of animals (rodents), which is often below the sensitivity of colorimetric methods;
– Study of a correlation between low serum UCB and different physiological conditions in humans;
– Evaluation of low UCB as a potential diagnostic marker;
– Cerebrospinal fluid analysis.
In cell/tissue culture growth medium:
– Oxidative stress response via activation of heme oxygenase-1;
– Rate of heme catabolism/turnover;
– Deviations in heme synthesis and intracellular transport;
– Red blood cells development, senescence, and heme recycling in in-vitro systems;
– Heme oxygenase activity;
– UCB transport;
– Antioxidant screening.
The assay is based on ligand-activated fluorescence. Fluorescence resulting from the non-covalent binding of UCB to the fluorescent agent is linearly proportional to the amount of UCB in the sample. Fluorescence can be measured using a standard GFP/FITC set of filters (optimal: 489 nm excitation/ 523 nm emission). The assay is specific for UCB, both albumin-bound and free. The assay is not suitable for conjugated bilirubin and other bilirubin derivatives.
Designed for use with 96-well plate systems. Can be adapted for use with cuvettes.