INFORMATION
| Full Name | Complement Component 4 |
|---|---|
| Synonym | C4 |
| Assay Type | Sandwich ELISA |
| Reactivity | Human |
| Range | 4.69-300ng/mL |
| Sensitivity | 2.81ng/mL |
| Sample Type | Serum, plasma and other biological fluids |
| Sample Volume | 100μL |
| Detection Wavelength | OD450 |
| Reaction Duration | 3.25h |
| Transportation Temperature | 2-8°C |
| Specificity | Specifically binds with C4, no obvious cross reaction with other analogues. |
| Microplate | 96-wells plate breakable into 12 x 8 wells strip |
ELISA KIT COMPONENTS
Upon receipt, unpack promptly and store as recommended in the instructions.
| Components | Specifications | Storage and Notes |
|---|---|---|
| Micro Plate | 96T: 8 wells×12 strips 48T: 8 wells×6 strips | Unopened: -20°C, 12 months Unused: Put it back in the aluminum foil bag and seal it, store it at -20°C. |
| Reference Standard | 96T: 2 vials 48T: 1 vial | Unopened: -20°C, 12 months Please use freshly dissolved standards for each experiment. Discard any unused standards after dissolution. |
| Biotinylated Detection Ab Concentrate (100×) | 96T: 120μL×1 vial 48T: 60μL×1 vial | Unopened: -20°C, 12 months Unused: Please seal the concentrate and store it at -20°C, and discard the working solution. |
| HRP Conjugate Concentrate (100×) | 96T: 120μL×1 vial 48T: 60μL×1 vial | Unopened: -20°C(Protect from light), 12 months Unused: Please seal the concentrate and store it at -20°C, and discard the working solution. |
| Biotinylated Detection Ab Diluent | 14mL×1 | 2-8℃, 12 months |
| HRP Conjugate Diluent | 14mL×1 | 2-8℃, 12 months |
| Reference Standard & Sample Diluent | 20mL×1 | 2-8℃, 12 months |
| Washing Buffer Concentrate (25×) | 30mL×1 | 2-8℃, 12 months |
| Substrate Reagent(TMB) | 10mL×1 | 2-8°C(Protect from light),12 months |
| Stop Solution | 7mL×1 | 2-8°C/Room temperature |
REQUIRED INSTRUMENTS AND REAGENTS
- Microplate reader (450nm wavelength filter).
- Incubator capable of maintaining 37℃.
- 1.5mL EP tube, absorbent paper.
- Pipette and disposable tip: 0.5-10μL, 2-20μL, 20-200μL, 200-1000μL.
- Double distilled water or deionized water.
ASSAY PROCEDURE SUMMARY
STANDARD CURVE
| STD.(ng/mL) | OD-1 | OD-2 | Average |
|---|---|---|---|
| 0 | 0.076 | 0.092 | 0.084 |
| 4.69 | 0.131 | 0.135 | 0.133 |
| 9.38 | 0.193 | 0.163 | 0.178 |
| 18.75 | 0.237 | 0.229 | 0.233 |
| 37.5 | 0.422 | 0.452 | 0.437 |
| 75 | 0.918 | 0.916 | 0.917 |
| 150 | 1.63 | 1.638 | 1.634 |
| 300 | 2.409 | 2.449 | 2.429 |
RECOVERY
5 different levels of target protein spiked in samples were evaluated for recovery experiments to determine the range of recovery and calculate the average recovery rate.
LINEARITY
5 different levels of target protein spiked in samples were evaluated for recovery experiments to determine the range of recovery and calculate the average recovery rate. Subsequently, the 5 samples were diluted to 2-fold, 4-fold, 8-fold, and 16-fold 10 concentrations, and recovery experiments were performed again to evaluate the range and average recovery rate at different dilution factors.
PRECISION(%)
Intra-assay precision (Precision within an assay): Low, mid range and high levels of the samples were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): Low, mid range and high levels of the samples were tested on 3 different plates, 20 replicates in each plate, respectively.
| Item | Intra-assay Precision | Inter-assay Precision | ||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (ng/mL) | 16.59 | 39.54 | 131.25 | 15.92 | 41.87 | 123.63 |
| Standard deviation | 0.86 | 1.88 | 5.37 | 0.84 | 1.89 | 6.53 |
| CV(%) | 5.16 | 4.76 | 4.09 | 5.29 | 4.51 | 5.28 |
STABILITY
Perform stability testing of the kit at 37°C and 2-8°C and obtain relevant data.
| ELISA kit(n=5) | 37°C for 1 month | 2-8°C for 6 months |
|---|---|---|
| Average(%) | 80 | 95-100 |
