Rat Dihydrotestosterone,DHT ELISA Kit
Product Details
| Description |
The Rat Dihydrotestosterone (DHT) ELISA Kit is designed for the quantitative determination of endogenic rat DHT concentrations in serum, plasma, or tissue homogenates. This assay employs the competitive inhibition enzyme immunoassay technique. The microtiter plate has been pre-coated with the goat-anti-rabbit DHT antibody. Standards or samples are added to the microwell plate with an anti-mouse DHT antibody and HRP-conjugate. The competitive inhibition reaction is launched between with HRP labeled DHT and unlabeled DHT (standard and sample) with the anti-mouse DHT antibody. These Ag-Ab complexes are captured by the antibody immobilized on the plate. Substrate A and B solution are mixed and then added to the wells, and the color turns blue. The color development is stopped after adding the stop solution, and the intensity of the color is in opposition to the amount of DHT in the sample. Read the OD value with the Microplate reader and calculated the DHT concentration referring to the st
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| Target Name |
Dihydrotestosterone,DHT |
| Alternative Names |
N/A |
| Abbreviation |
DHT |
| Species |
Rattus norvegicus (Rat) |
| Sample Types |
serum, plasma, tissue homogenates |
| Detection Range |
10 pg/mL-2000 pg/mL |
| Sensitivity |
5 pg/mL |
| Assay Time |
1-5h |
| Sample Volume |
50-100ul |
| Detection Wavelength |
450 nm |
| Research Area |
Signal Transduction |
| Assay Principle |
quantitative |
| Measurement |
Competitive |
| Precision |
| Intra-assay Precision (Precision within an assay): CV% |
| 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% |
| 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 DHT in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay. |
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Sample |
Serum(n=4) |
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| 1:1 |
Average % |
102 |
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| Range % |
98-106 |
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| 1:2 |
Average % |
93 |
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| Range % |
89-97 |
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| 1:4 |
Average % |
95 |
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| Range % |
89-99 |
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| 1:8 |
Average % |
102 |
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| Range % |
98-106 |
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| Recovery |
| The recovery of rat DHT 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. |
| Sample Type |
Average % Recovery |
Range |
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| Serum (n=5) |
92 |
87-95 |
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| EDTA plasma (n=4) |
102 |
97-106 |
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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. |
| pg/ml |
OD1 |
OD2 |
Average |
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| 2000 |
0.156 |
0.160 |
0.158 |
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| 500 |
0.312 |
0.329 |
0.321 |
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| 160 |
0.498 |
0.528 |
0.513 |
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| 40 |
0.818 |
0.787 |
0.803 |
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| 10 |
1.316 |
1.278 |
1.297 |
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| 0 |
1.960 |
1.954 |
1.957 |
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| Materials provided |
- A 96-well Assay plate -- The 96-well plate has been pre-coated with goat-anti-rabbit DHT antibody.
- Standard 6 x 0.5 ml -- Dilute the standard at dilution series, read the OD values, and then draw a standard curve.
- Anti-mouse DHT Antibody 1 x 6 ml -- Act as the detection antibody.
- HRP-conjugate 1 x 6 ml -- Bind to the DHT antibody, and HRP catalyzes the TMB 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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| Storage |
Store at 2-8°C. Please refer to protocol. |
| Lead Time |
3-5 working days |
