Background

Leukotrienes C4/ D4/ E4 (LTC4/D4/E4) are potent mediators of immediate hypersensitivity, bronchoconstriction, smooth muscle contraction, increased vascular permeability, and epithelial mucous secretion. They are derived from arachidonic acid through the 5-lipoxygenase pathway. The monoclonal antibody in this kit recognizes LTC4, LTD4 and LTE4 and allows for the quantitation of all of these potent leukotrienes in a single assay. This ELISA provides quantitative analysis of LTC4/D4/E4 levels in biological fluids.

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Assay Principle

This is an ELISA? for the quantitative analysis of Leukotriene C4/D4/E4 levels in biological fluid. This test kit operates on the basis of competition between the enzyme conjugate and Leukotriene C4/D4/E4 in the sample for a limited number of binding sites.

First, the sample or standard solution is added to the microplate.? Next, the diluted enzyme conjugate is added and the mixture is shaken and incubated at room temperature for one hour. During the incubation, competition for binding sites is taking place. The plate is then washed, removing all the unbound material. The bound enzyme conjugate is detected by the addition of substrate which generates an optimal color after 30 minutes. Quantitative test results may be obtained by measuring and comparing the absorbance reading of the wells of the samples against the standards with a microplate reader at 450nm or 650nm. The extent of color development is inversely proportional to the amount of Leukotriene C4/D4/E4 in the sample or standard.? For example, the absence of Leukotriene C4/D4/E4_ in the sample will result in a bright blue color, whereas the presence of Leukotriene C4/D4/E4 will result in decreased or no color development.

If using urine as a sample it is recommended to run our

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Creatinine Assay kit (CR01)

in conjunction with your samples since urinary creatinine levels may be used to normalize the rate of excretion of other analytes.

Cross Reactivity

??? Leukotriene C4 ??? ??? ??? ??? ??? 100.0%
??? Leukotriene D4??? ??? ??? ??? ??? ?? 80.0%
??? Leukotriene? E4??? ??? ??? ??? ??? ? 80.0%
??? Leukotriene? A4??? ??? ??? ??? ??? ? 2.00%
??? Leukotriene? B4??? ??? ??? ??? ??? <1.00%
??? Prostaglandin? B2??? ??? ??? ??? <0.01%
??? Prostaglandin D2??? ??? ??? ???? <0.01%
??? Prostaglandin? E2??? ??? ??? ??? <0.01%
??? Prostaglandin? F2α??? ??? ??? ? <0.01%
??? 6-keto-Prostaglandin? F1α? <0.01%
??? 5-HETE??? ??? ??? ??? ??? ??? ??? ??? ? <0.01%
??? 12-HETE??? ??? ??? ??? ??? ??? ??? ??? <0.01%
??? 15-HETE??? ??? ??? ??? ??? ??? ??? ??? <0.01%
??? 6-trans-Leukotriene B4??? ??? <0.01%
??? 20-OH-Leukotriene B4??? ??? <0.01%
??? Thromboxane? B2??? ??? ??? ??? <0.01%

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