INFORMATION
| Full Name | Big Dynorphin |
|---|---|
| Synonym | Dyn |
| Assay Type | Competitive ELISA |
| Reactivity | Guinea pig |
| Range | 7.81-500pg/mL |
| Sensitivity | 4.69pg/mL |
| Sample Type | Serum, plasma and other biological fluids |
| Sample Volume | 50μL |
| Detection Wavelength | OD450 |
| Reaction Duration | 1.5h |
| Transportation Temperature | 2-8°C |
| Specificity | Specifically binds with Dyn, 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.(pg/mL) | OD-1 | OD-2 | Average |
|---|---|---|---|
| 0 | 1.961 | 1.961 | 1.961 |
| 7.82 | 1.639 | 1.641 | 1.64 |
| 15.63 | 1.384 | 1.374 | 1.379 |
| 31.25 | 1.059 | 1.029 | 1.044 |
| 62.5 | 0.712 | 0.736 | 0.724 |
| 125 | 0.509 | 0.481 | 0.495 |
| 250 | 0.339 | 0.385 | 0.362 |
| 500 | 0.292 | 0.296 | 0.294 |
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 (pg/mL) | 27.5 | 58.3 | 221.84 | 28.38 | 54.93 | 216.02 |
| Standard deviation | 1.68 | 3.47 | 9.49 | 1.65 | 2.42 | 11.1 |
| CV(%) | 6.11 | 5.96 | 4.28 | 5.8 | 4.41 | 5.14 |
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 |
