Rat D2D(D-Dimer) ELISA Kit
SKU: E-EL-R0317
Rat D2D(D-Dimer) ELISA Kit
Detection Range | 78.13-5000 ng/mL |
Sensitivity | 46.88 ng/mL |
Product Details
Properties
Format | 96T |
Assay time | 2.0h |
Sample type | plasma |
Sample volume | 50μL |
Sample size | Run up to 46 samples in duplicate and 30 samples in triplicate |
Storage | 2-8℃ |
Interpretation | Detection range: 78.13-5000 ng/mL Sensitivity: 46.88 ng/mL |
Application | Quantitative determination of Human concentrations in human serum, plasma and other biological fluids |
Reproducibility | Both intra-CV and inter-CV are < 10% |
Test Principle
This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Rat D2D. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Rat D2D and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Rat D2D, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Rat D2D. You can calculate the concentration of Rat D2D in the samples by comparing the OD of the samples to the standard curve.
Kit Components and Storage
An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.
Item | Specifications | Storage |
---|---|---|
Micro ELISA Plate(Dismountable) | 96T: 8 wells ×12 strips 48T: 8 wells ×6 strips |
-20℃, 6 months |
Reference Standard | 96T: 2 vials 48T: 1 vial |
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Concentrated Biotinylated Detection Ab (100×) | 96T: 1 vial, 120 μL 48T: 1 vial, 60 μL |
|
Concentrated HRP Conjugate (100×) | 96T: 1 vial, 120 μL 48T: 1 vial, 60 μL |
-20℃(Protect from light), 6 months |
Reference Standard & Sample Diluent | 1 vial, 20 mL | 2-8°C, 6 months |
Biotinylated Detection Ab Diluent | 1 vial, 14 mL | |
HRP Conjugate Diluent | 1 vial, 14 mL | |
Concentrated Wash Buffer (25×) | 1 vial, 30 mL | |
Substrate Reagent | 1 vial, 10 mL | 2-8℃(Protect from light) |
Stop Solution | 1 vial, 10 mL | 2-8°C |
Plate Sealer | 5 pieces | |
Manual | 1 copy | |
Certificate of Analysis | 1 copy |
Technical Data:
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and
high level Rat D2D were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and
high level Rat D2D were tested on 3 different plates, 20 replicates in each plate,
respectively.
Intra-assay Precision | Inter-assay Precision | |||||
---|---|---|---|---|---|---|
Sample | 1 | 2 | 3 | 1 | 2 | 3 |
n | 20 | 20 | 20 | 20 | 20 | 20 |
Mean (ng/mL) | 250.9 | 499.7 | 2441.3 | 265.8 | 545.6 | 2642.9 |
Standard deviation | 17.3 | 21 | 119.6 | 13.6 | 31.6 | 105.7 |
CV (%) | 6.9 | 4.2 | 4.9 | 5.12 | 5.79 | 4 |
Recovery
The recovery of Rat D2D spiked at three different levels in samples throughout the
range of the assay was evaluated in various matrices.
Sample Type | Range (%) | Average Recovery (%) |
---|---|---|
EDTA plasma (n=8) | 88-100 | 94 |
Assay Procedures
1. Add 50μL standard or sample to the wells, immediately add 50μL Biotinylated Detection Ab working solution to each well. Incubate for 45 min at 37°C |
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2. Aspirate and wash the plate for 3 times |
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3. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times |
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4. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C |
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5. Add 50μL Stop Solution |
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6. Read the plate at 450nm immediately. Calculation of the results |