Background

Thromboxane B2 (TXB2) is a stable hydrolysis product of Thromboxane A2 (TXA2), which is derived from Prostaglandin H2 (PGH2) by the action of thromboxane synthase. It is a major product following platelet aggregation induced by a variety of agents such as thrombin and collagen. This competitive ELISA permits quantitative analysis of TXB2 levels in biological fluids. Usually, urine and tissue culture supernatant can be assayed directly, whereas TXB2 should be extracted from plasma prior to analysis.
 

Assay Principle

This is an ELISA for the quantitative analysis of TXB2 levels in biological fluid.  This test kit operates on the basis of competition between the enzyme conjugate and the 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 TXB2 in the sample or standard.  For example, the absence of TXB2 in the sample will result in a bright blue color, whereas the presence of 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.

Cross Reactivity

Thromboxane B2                                              100.0%
2,3-dinor-Thromboxane B2                                 30.0%
Prostaglandin D                                                21.21%
Prostaglandin E                                                20.08%
11-dehydro-Thromboxane B                              20.07%
Prostaglandin F2a                                              0.06%
6-keto-Prostaglandin F1a                                    0.05%
Prostaglandin F1a                                               0.02%
Arachidonic Acid                                               <0.01%
Leukotriene B4                                                 <0.01%
Prostaglandin A2                                              <0.01%
Prostaglandin B2                                              <0.01%
13,14-dihydro-15-keto-Prostaglandin F2a           <0.01%