INFORMATION
| Full Name | Interleukin-2 Receptor alpha chain |
|---|---|
| Synonym | Interleukin-2 receptor subunit alpha, IL2RA, IL-2 receptor subunit alpha, IL-2-RA, IL-2R subunit alpha, IL2-RA, TAC antigen, p55, CD25 |
| Assay Type | Sandwich |
| Reactivity | Rat |
| Range | 78.13-5000pg/mL |
| Sensitivity | 46.88pg/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 IL-2Rα/CD25, 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 | 0.069 | 0.079 | 0.074 |
| 78.13 | 0.126 | 0.134 | 0.13 |
| 156.25 | 0.199 | 0.169 | 0.184 |
| 312.5 | 0.308 | 0.3 | 0.304 |
| 625 | 0.491 | 0.491 | 0.491 |
| 1250 | 1.052 | 1.04 | 1.046 |
| 2500 | 1.717 | 1.735 | 1.726 |
| 5000 | 2.521 | 2.565 | 2.543 |
RECOVERY
The recovery of Rat IL-2Rα/CD25 spiked to three different levels in samples throughout the range of the assay in various matrices was evaluated.
LINEARITY
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of Rat IL-2Rα/CD25 in various matrices were diluted with the Reference Standard & Sample Diluent to produce samples with values within the dynamic range of the assay.
PRECISION(%)
Intra-Assay Precision (Precision within an assay): Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays): Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components.
| Item | Intra-assay Precision | Inter-assay Precision | ||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 247.27 | 599.12 | 2292.86 | 261.68 | 583.14 | 2308.26 |
| Standard deviation | 15.11 | 30.08 | 75.66 | 15.62 | 28.52 | 124.65 |
| CV(%) | 6.11 | 5.02 | 3.3 | 5.97 | 4.89 | 5.4 |
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 |
