Intended use
This immunoassay kit allows for the in vitro quantitative determination of General FK506 concentrations in serum, Plasma, tissue homogenates and Cell culture supernates and Other biological fluids.
Test principle
The ELISA is based on the competitive binding enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with an antibody specific to General FK506, During the reaction, General FK506 in the sample or standard competes with a fixed amount of biotin-labeled General FK506 for sites on a pre-coated Monoclonal antibody specific to General FK506. Excess conjugate and unbound sample or standard are washed from the plate. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of General FK506 in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Precision

Intra-assay Precision (Precision within an assay):Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.

Intra-Assay CV: ≤3.9%

Inter-assay Precision (Precision between assays):Three samples of known concentration were tested in five separate assays to assess inter-assay precision.

Inter-Assay CV: ≤7.7%

Recovery

Recovery was determined by spiking various levels of Thromboxane A2 into serum and plasma.

Sample Type	Average(%)	Recovery Range(%)
Serum	99	91-103
Plasma	98	93-106

 

 

 

 

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Thromboxane A2 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample	1：2	1:4	1:8	1:16
serum(n=5)	90-100%	90-102%	94-103%	94-102%
EDTA plasma(n=5)	83-95%	90-99%	86-96%	107-117%
heparin plasma(n=5)	110-119%	99-110%	109-119%	91-102%

 

General Annotation

Function:

Thromboxane A2 (TXA2) is a thromboxane. It is produced by activated platelets and has prothrombotic properties: it stimulates activation of new platelets as well as increases platelet aggregation. This is achieved by mediating expression of the glycoprotein complex GP IIb/IIIa in the cell membrane of platelets. Circulating fibrinogen binds these receptors on adjacent platelets, further strengthening the clot.

 

Location:

TXA2 is generated from prostaglandin H2 by thromboxane-A synthase. Aspirin irreversibly inhibits platelet cyclooxygenase 1 preventing the formation of prostaglandin H2, and therefore thromboxane A2.

 

Sequence:C20H32O5

 

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