Leukotriene B4 (LTB4) is the most potent chemotactic substance ever described for leukocytes. Using this competitive ELISA, LTB4 in urine and tissue culture supernatants can be assayed directly, whereas LTB4 should be extracted from plasma prior to analysis.
This is an ELISA for the quantitative analysis of LTB4 levels in biological fluid. This test kit operates on the basis of competition between the enzyme conjugate and the LTB4 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 LTB4 in the sample or standard. For example, the absence of LTB4 in the sample will result in a bright blue color, whereas the presence of LTB4 will result in decreased or no color development.
If using urine as a sample it is recommended to run our
in conjunction with your samples since urinary creatinine levels may be used to normalize the rate of excretion of other analytes.