Background

Measurement of 11-Dehdydrothromboxane B2 (11-d-TXB2), one of the major metabolites of Thromboxane B2 (TXB2), in plasma is an approach to the assessment of thromboxane formation in circulation. Usually, urine and tissue culture supernatants can be assayed directly using this competitive ELISA, whereas 11-d-TXB2 should be extracted from plasma prior to analysis.

Assay Principle

This is an ELISA  for the quantitative analysis of 11-d-TXB2 levels in biological fluid.  This test kit operates on the basis of competition between the enzyme conjugate and the 11-d-TXB2 in the sample for a limited number of binding sites on the antibody coated plate.

The sample or standard solution is first 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 11-d-TXB2 in the sample or standard.  For example, the absence of 11-d-TXB2  in the sample will result in a bright blue color, whereas the presence of 11-d-TXB2 will result in decreased or no color development.

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

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.