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| Organism species | Rattus norvegicus (Rat) |
| Product No. | YBG433Ra |
| Sample type | Tissue homogenates and other biological fluids. |
| Format | 96T |
| Assay length | 4.5 hours |
| Detection range | 0.781-50ng/mL The standard curve concentrations used for the ELISA’s were 50ng/mL, 25ng/mL, 12.5ng/mL, 6.25ng/mL, 3.125ng/mL, 1.563ng/mL, 0.781ng/mL |
| Sensitivity | The minimum detectable dose of this kit is typically less than 0.24ng/mL. |
No significant cross-reactivity or interference between Cadherin EGF LAG Seven Pass G-Type Receptor 2 (CELSR2) and analogues was observed.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Cadherin EGF LAG Seven Pass G-Type Receptor 2 (CELSR2) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.
| Reagents | Quantity | Reagents | Quantity |
| Pre-coated, ready to use 96-well strip plate | 1 | Plate sealer for 96 wells | 4 |
| Standard | 2 | Standard Diluent | 1×20mL |
| Detection Reagent A | 1×120µL | Assay Diluent A | 1×12mL |
| Detection Reagent B | 1×120µL | Assay Diluent B | 1×12mL |
| TMB Substrate | 1×9mL | Stop Solution | 1×6mL |
| Wash Buffer (30 × concentrate) | 1×20mL | Instruction manual | 1 |
2. Add 100µL standard or sample to each well. Incubate 2 hours at 37oC;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37oC;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37oC;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 15-25 minutes at 37oC;
8. Add 50µL Stop Solution. Read at 450nm immediately.
The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Cadherin EGF LAG Seven Pass G-Type Receptor 2 (CELSR2). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Cadherin EGF LAG Seven Pass G-Type Receptor 2 (CELSR2). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Cadherin EGF LAG Seven Pass G-Type Receptor 2 (CELSR2), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Cadherin EGF LAG Seven Pass G-Type Receptor 2 (CELSR2) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
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