| Description |
This Rat 5HT ELISA Kit was designed for the quantitative measurement of Rat 5HT protein in serum, plasma, cell lysates, tissue homogenates. It is a Competitive ELISA kit, its detection range is 0.8 ng/mL-160 ng/mL and the sensitivity is 0.4 ng/mL. |
| Target Name |
5-Hydroxytryptamine,5HT |
| Abbreviation |
5HT |
| Species |
Rattus norvegicus (Rat) |
| Sample Types |
serum, plasma, cell lysates, tissue homogenates |
| Detection Range |
0.8 ng/mL-160 ng/mL |
| Sensitivity |
0.4 ng/mL |
| Assay Time |
1-5h |
| Sample Volume |
50-100ul |
| Detection Wavelength |
450 nm |
| Research Area |
Neuroscience
|
| Assay Principle |
quantitative |
| Measurement |
Competitive |
| Precision |
| Intra-assay Precision (Precision within an assay): CV%<15% |
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| Three samples of known concentration were tested twenty times on one plate to assess. |
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| Inter-assay Precision (Precision between assays): CV%<15% |
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| Three samples of known concentration were tested in twenty assays to assess. |
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| Linearity |
| To assess the linearity of the assay, samples were spiked with high concentrations of rat 5-HT in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay. |
| |
Sample |
Serum(n=4) |
|
| 1:1 |
Average % |
94 |
|
| Range % |
89-98 |
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| 1:2 |
Average % |
95 |
|
| Range % |
92-99 |
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| 1:4 |
Average % |
97 |
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| Range % |
89-103 |
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| 1:8 |
Average % |
95 |
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| Range % |
90-100 |
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| Recovery |
| The recovery of rat 5-HT spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section. |
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| Sample Type |
Average % Recovery |
Range |
|
| Serum (n=5) |
96 |
91-102 |
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| EDTA plasma (n=4) |
96 |
92-103 |
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| Typical Data |
| These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed. |
|
| ng/ml |
OD1 |
OD2 |
Average |
|
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| 160 |
0.069 |
0.074 |
0.072 |
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| 40 |
0.181 |
0.195 |
0.188 |
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| 12.8 |
0.426 |
0.439 |
0.433 |
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| 3.2 |
0.802 |
0.855 |
0.829 |
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| 0.8 |
1.155 |
1.231 |
1.193 |
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| 0 |
2.158 |
2.227 |
2.193 |
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| Materials provided |
- A 96-well Assay plate -- The 96-well plate has been pre-coated with goat-anti-rabbit 5HT antibody.
- Standard 6 x 0.5 ml -- Dilute the standard at dilution series, read the OD values, and then draw a standard curve.
- Anti-rat 5HT Antibody 1 x 6 ml -- Act as the detection antibody.
- HRP-conjugated 5HT 1 x 6 ml -- Bind to the 5HT antibody, and HRP catalyzes the substrate A and B to elicit a chromogenic reaction.
- Wash Buffer (20 x concentrate) 1 x 15 ml -- Wash away unbound or free substances.
- Substrate A 1 x 7 ml -- Mix with substrate B, and the TMB is catalyzed by HRP to develop color.
- Substrate B 1 x 7 ml -- Mix with substrate A, and the TMB is catalyzed by HRP to develop color.
- Stop Solution 1 x 7 ml -- Stop the color reaction. The solution color immediately turns from blue to yellow.
- Four Adhesive Strips (For 96 wells)
- An Instruction manual
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| Materials not provided |
- A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 600 nm - 630 nm.
- An incubator that can provide stable incubation conditions up to 37°C±5°C.
- Centrifuge
- Vortex
- Squirt bottle, manifold dispenser, or automated microplate washer
- Absorbent paper for blotting the microtiter plate
- 50-300ul multi-channel micropipette
- Pipette tips
- Single-channel micropipette with different ranges
- 100ml and 500ml graduated cylinders
- Deionized or distilled water
- Timer
- Test tubes for dilution
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| ELISA Data Analysis |
Watch ELISA data processing video & download Curve Expert if needed |
Troubleshooting
and FAQs |
ELISA kit FAQs |
| Storage |
Store at 2-8°C. Please refer to protocol. |
| Lead Time |
3-5 working days |
