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Porcine sC5b-9(Soluble Terminal Complement Complex C5b-9) ELISA Kit
SKU: E-EL-P0485
Porcine sC5b-9(Soluble Terminal Complement Complex C5b-9) ELISA Kit
| Detection Range | 7.81-500 ng/mL |
| Sensitivity | 4.69 ng/mL |
Product Details
Properties
| Assay type | Sandwich-ELISA |
| Format | 96T |
| Assay time | 3.5h |
| Detection range | 7.81-500 ng/mL |
| Sensitivity | 4.69 ng/mL |
| Sample type &Sample volume | Serum, plasma and other biological fluids; 100μL |
| Specificity | This kit recognizes Porcine sC5b-9 in samples. No significant cross-reactivity or interference between Porcine sC5b-9 and analogues was observed |
| Reproducibility | Coefficient of variation is < 10% |
| Application | This ELISA kit applies to the in vitro quantitative determination of Porcine sC5b-9 concentrations . |
Test Principle
This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in
this kit has been pre-coated with an antibody specific to Porcine sC5b-9. Samples (or
Standards) are added to the micro ELISA plate wells and combined with the specific
antibody. Then a biotinylated detection antibody specific for Porcine sC5b-9 and AvidinHorseradish 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 Porcine sC5b-9, 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 ± 2 nm. The OD value is proportional to the concentration of Porcine sC5b-9. You can calculate the concentration of Porcine sC5b-9 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 |
|
| 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 Porcine sC5b-9 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and
high level Porcine sC5b-9 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 (pg/mL) | 25 | 62.68 | 229.72 | 26.67 | 66.84 | 241.42 |
| Standard deviation | 1.53 | 3.54 | 7.51 | 1.49 | 3.63 | 7.34 |
| CV (%) | 6.12 | 5.65 | 3.27 | 5.59 | 5.43 | 3.04 |
Recovery
The recovery of Porcine sC5b-9 spiked at three different levels in samples throughout
the range of the assay was evaluated in various matrices.
| Sample Type | Range (%) | Average Recovery (%) |
|---|---|---|
| Serum(n=8) | 87-101 | 93 |
| EDTA plasma (n=8) | 87-99 | 93 |
| Cell culture media (n=8) | 84-96 | 90 |
Assay Procedures
 |
1. Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C |
 |
2. Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C |
 |
3. Aspirate and wash the plate for 3 times |
 |
4. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times |
 |
5. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C |
 |
6. Add 50μL Stop Solution |
 |
7. Read the plate at 450nm immediately. Calculation of the results |