In this assay CysLT present in sample competes with the fixed amount of CysLT conjugated to alkaline phosphatase for the limited number of binding sites on the monoclonal anti-CysLT antibody. The antigen-antibody complex binds to the monoclonal anti-mouse IgG antibody-coated multiwell plate. A substrate is added which reacts with the bound enzyme. The reaction is stopped and absorbance read at 405 nm.The intensity of the yellow color read is inversely proportional to the concentration of CysLT in the standards or the samples.
Cysteinyl leukotriene (CysLT) causes smooth muscle contraction and increased capillary permeability. Detection of CysLT levels is useful in hepatitis B, Helicobacter pylori infections and in cerebral vasospasms.
Features and Benefits
The assay is specific for CysLT and shows negligible cross-reactivity with 20-25 related eicosanoid compounds tested (from <0.1 to 2%). It is quantitative, with a sensitivity of 26 pg/mL. Precoated plates eliminate the coating step. All incubations proceed at room temperature. Any commercially available microwell plate reader may be used. Results are available in 4 hours and recovery ranges from 99-111%.