| Description |
The copeptin ELISA kit is used to detect and quantify protein levels of endogenous copeptin. The assay recognizes human copeptin in serum, urine, cell culture supernates, and tissue homogenates. It provides a detection range of 78 pg/mL-5000 pg/mL. It has a high sensitivity, as low as 19.5 pg/mL. In addition, this kit also possesses high precision, good lineaity, excellent specificity, and high recovery. This experiment uses the sandwich ELISA technique. The pre-coated antibody on the micro ELISA plate is an anti-human copeptin antibody and the detection antibody is a biotin-conjugated polyclonal copeptin antibody. Samples and biotin labeling antibody are added into wells and thoroughly washed out. Avidin-HRP conjugates are added to wells. The TMB substrate is added for coloring after thorough washing. The solution turns blue in HRP catalytic and finally turns yellow under the action of the stop solution. The color depth is positively correlated to the testing analyte in samples. Copep.
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| Target Name |
copeptin |
| Abbreviation |
copeptin |
| Species |
Homo sapiens (Human) |
| Sample Types |
serum, urine, cell culture supernates, tissue homogenates |
| Detection Range |
78 pg/mL-5000 pg/mL |
| Sensitivity |
19.5 pg/mL |
| Assay Time |
1-5h |
| Sample Volume |
50-100ul |
| Detection Wavelength |
450 nm |
| Research Area |
Others |
| Assay Principle |
quantitative |
| Measurement |
Sandwich |
| 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 human Copeptin 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 % |
89 |
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| Range % |
82-95 |
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| 1:2 |
Average % |
99 |
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| Range % |
92-105 |
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| 1:4 |
Average % |
97 |
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| Range % |
90-106 |
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| 1:8 |
Average % |
96 |
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| Range % |
91-102 |
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| Recovery |
| The recovery of human Copeptin 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) |
93 |
88-98 |
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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 |
Corrected |
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| 5000 |
2.535 |
2.752 |
2.644 |
2.533 |
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| 2500 |
2.012 |
2.179 |
2.096 |
1.985 |
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| 1250 |
1.598 |
1.599 |
1.599 |
1.488 |
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| 625 |
0.847 |
0.867 |
0.857 |
0.746 |
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| 312 |
0.478 |
0.502 |
0.490 |
0.379 |
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| 156 |
0.336 |
0.356 |
0.346 |
0.235 |
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| 78 |
0.237 |
0.221 |
0.229 |
0.118 |
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| 0 |
0.109 |
0.112 |
0.111 |
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| Materials provided |
- A micro ELISA plate ---The 96-well plate has been pre-coated with an anti-Human copeptin antibody.
- Two vials lyophilized standard ---Dilute a bottle of the standard at dilution series, read the OD values, and then draw a standard curve.
- One vial Biotin-labeled copeptin antibody (100 x concentrate) (120 μl/bottle) ---Act as the detection antibody.
- One vial HRP-avidin (100 x concentrate) (120 μl/bottle) ---Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
- One vial Biotin-antibody Diluent (15 ml/bottle) ---Dilute the Biotin-antibody.
- One vial HRP-avidin Diluent (15 ml/bottle) ---Dilute the HRP-avidin solution.
- One vial Sample Diluent (50 ml/bottle) ---Dilute the sample to an appropriate concentration.
- One vial Wash Buffer (25 x concentrate) (20 ml/bottle) ---Wash away unbound or free substances.
- One vial TMB Substrate (10 ml/bottle) ---Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
- One vial Stop Solution (10 ml/bottle) ---Stop the color reaction. The solution color immediately turns from blue to yellow.
- Four Adhesive Strips (For 96 wells) ---Cover the microplate when incubating.
- 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 540 nm or 570 nm.
- An incubator can provide stable incubation conditions up to 37°C±5°
- 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 |
