KEY FEATURES
| Full Name | Serotonin/5-Hydroxytryptamine |
|---|---|
| Synonym | 5HT;Enteramine;Thrombocytin;Thrombotonin;3-(β-Aminoethyl)-5-hydroxyindole; |
| Assay Type | Competitive |
| Reactivity | Universal |
| Range | 15.63-1000ng/mL |
| Sensitivity | 9.38ng/mL |
| Sample Type | Serum,plasma and other biological fluids |
| Sample Volume | 50μL |
| Detection Wavelength | OD450 |
| Transportation Temperature | 2-8°C |
| Specificity | The kit detected ST/5-HT in the samples and no significant cross-species reactions were found |
| Microplate | 96-wells plate breakable into 12 x 8 wells strip |
TEST PRINCIPLE
| This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with ST/5-HT. During the reaction, ST/5-HT in the sample or standard competes with a fixed amount of ST/5-HT on the solid phase supporter for sites on the Biotinylated Detection Ab specific to ST/5-HT. Excess conjugate and unbound sample or standard are washed away, and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns from blue to yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of ST/5-HT in tested samples can be calculated by comparing the OD of the samples to the standard curve. |
ELISA KIT COMPONENTS
Upon receipt, unpack promptly and store as recommended in the instructions.
| Components | Specifications | Storage and Notes |
|---|---|---|
| Micro Plate | 96T: 8 wells×12 strips 48T: 8 wells×6 strips | Unopened: -20°C, 12 months Unused: Put it back in the aluminum foil bag and seal it, store it at -20°C. |
| Reference Standard | 96T: 2 vials 48T: 1 vial | Unopened: -20°C, 12 months Please use freshly dissolved standards for each experiment. Discard any unused standards after dissolution. |
| Biotinylated Detection Ab Concentrate (100×) | 96T: 120μL×1 vial 48T: 60μL×1 vial | Unopened: -20°C, 12 months Unused: Please seal the concentrate and store it at -20°C, and discard the working solution. |
| HRP Conjugate Concentrate (100×) | 96T: 120μL×1 vial 48T: 60μL×1 vial | Unopened: -20°C(Protect from light), 12 months Unused: Please seal the concentrate and store it at -20°C, and discard the working solution. |
| Biotinylated Detection Ab Diluent | 14mL×1 | 2-8℃, 12 months |
| HRP Conjugate Diluent | 14mL×1 | 2-8℃, 12 months |
| Reference Standard & Sample Diluent | 20mL×1 | 2-8℃, 12 months |
| Washing Buffer Concentrate (25×) | 30mL×1 | 2-8℃, 12 months |
| Substrate Reagent(TMB) | 10mL×1 | 2-8°C(Protect from light),12 months |
| Stop Solution | 7mL×1 | 2-8°C/Room temperature |
ASSAY PROCEDURES
TYPICAL DATA
ST/5-HT ELISA Standard Curve
Typical data are for reference only and curves should be replotted for each experiment. The Logistics function is recommended for fitting.
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.
Inter-Assay Precision (Precision between assays): Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components.
| Intra-assay Precision | Inter-assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (ng/mL) | 54.22 | 119.39 | 369.61 | 54.14 | 128.11 | 403.79 |
| Standard deviation | 3.69 | 7.09 | 12.42 | 3.7 | 6.32 | 16.52 |
| CV(%) | 6.81 | 5.94 | 3.36 | 6.83 | 4.93 | 4.09 |
RECOVERY
The recovery of ST/5-HT spiked to three different levels in samples throughout the range of the assay in various matrices was evaluated.
| Sample Type | Range (%) | Average Recovery (%) |
|---|---|---|
| Serum(n=8) | 88-103 | 95 |
| EDTA plasma (n=8) | 88-99 | 94 |
| Cell culture media (n=8) | 91-107 | 98 |
LINEARITY
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of ST/5-HT in various matrices were diluted with the Reference Standard & Sample Diluent to produce samples with values within the dynamic range of the assay.
