Intended use 
This immunoassay kit allows for the in vitro quantitative determination of General 5HT 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 5HT, During the reaction, General 5HT in the sample or standard competes with a fixed amount of biotin-labeled General 5HT for sites on a pre-coated Monoclonal antibody specific to General. 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 5HT 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: ≤4.7%

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 Serotonin into serum and plasma.

Sample Type	Average(%)	Recovery Range(%)
Serum	97	91-103
Plasma	99	93-105

 

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Serotonin 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)	98-107%	107-117%	90-100%	83-92%
EDTA plasma(n=5)	95-105%	113-123%	93-103%	101-113%
heparin plasma(n=5)	106-118%	102-112%	105-117%	95-104%

 

General Annotation

 

Function:

Serotonin or 5-hydroxytryptamine (5-HT) is a monoamine neurotransmitter. Biochemically derived from tryptophan, serotonin is primarily found in the gastrointestinal (GI) tract, platelets, and in the central nervous system (CNS) of animals including humans. It is popularly thought to be a contributor to feelings of well-being and happiness.

 

Location:

Approximately 90% of the human body's total serotonin is located in the enterochromaffin cells in the gut, where it is used to regulate intestinal movements. The remainder is synthesized in serotonergic neurons of the CNS where it has various functions. These include the regulation of mood, appetite, and sleep. Serotonin also has some cognitive functions, including memory and learning. Modulation of serotonin at synapses is thought to be a major action of several classes of pharmacological antidepressants.

 

Sequence:C10H12N2O

 

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