INFORMATION
| Full Name | cyclic guanosine monophosphate |
|---|---|
| Synonym | cGMP |
| Assay Type | Competitive |
| Reactivity | Rabbit |
| Range | 1.56-100ng/mL |
| Sensitivity | 0.94ng/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 cGMP, 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 | 2.162 | 2.172 | 2.167 |
| 1.57 | 1.865 | 1.869 | 1.867 |
| 3.13 | 1.629 | 1.623 | 1.626 |
| 6.25 | 1.311 | 1.287 | 1.299 |
| 12.5 | 0.948 | 0.958 | 0.953 |
| 25 | 0.684 | 0.66 | 0.672 |
| 50 | 0.488 | 0.49 | 0.489 |
| 100 | 0.385 | 0.387 | 0.386 |
RECOVERY
The recovery of Rabbit cGMP 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 Rabbit cGMP 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 (ng/mL) | 4.73 | 12.09 | 44.84 | 4.43 | 11.96 | 48.8 |
| Standard deviation | 0.32 | 0.51 | 2.18 | 0.24 | 0.62 | 2.59 |
| CV(%) | 6.74 | 4.22 | 4.87 | 5.33 | 5.19 | 5.3 |
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 |
