Description
| Product name: | Human PDE5A(Phosphodiesterase 5A, cGMP Specific) ELISA Kit |
| Reactivity: | Human |
| Alternative Names: | CGB-PDE; cGMP-binding cGMP-specific phosphodiesterase; cGMP-specific 3′,5′-cyclic phosphodiesterase |
| Assay Type: | Sandwich |
| Sensitivity: | 0.56 ng/mL |
| Standard: | 100 ng/mL |
| Range: | 1.57-100 ng/mL |
| Sample Type: | serum, plasma, tissue homogenates and other biological fluids |
| Assay Length: | 3.5h |
| Research Area: | Enzyme & Kinase; |
| Test principle: | The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Phosphodiesterase 5A, cGMP Specific(PDE5A). Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Phosphodiesterase 5A, cGMP Specific(PDE5A). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Phosphodiesterase 5A, cGMP Specific(PDE5A), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Phosphodiesterase 5A, cGMP Specific(PDE5A) in the samples is then determined by comparing the OD of the samples to the standard curve. |
| Standard Curve | 
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Precision
Intra-assay Precision (Precision within an assay)：CV%<8%
Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-assay Precision (Precision between assays)：CV%<10%
Three samples of known concentration were tested in forty separate assays to assess inter-assay precision.
Recovery
Matrices listed below were spiked with certain level of recombinant PDE5A and the recovery rates were calculated by comparing the measured value to the expected amount of PDE5A in samples.
| Matrix | Recovery range | Average |
|---|---|---|
| serum(n=5) | 81-93% | 87% |
| EDTA plasma(n=5) | 83-101% | 87% |
| Heparin plasma(n=5) | 86-94% | 90% |
Linearity
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of PDE5A and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
| Matrix | 1:2 | 1:4 | 1:8 | 1:16 |
|---|---|---|---|---|
| serum(n=5) | 79-93% | 89-97% | 85-94% | 87-101% |
| EDTA plasma(n=5) | 86-95% | 93-103% | 87-98% | 85-96% |
| Heparin plasma(n=5) | 86-97% | 93-105% | 82-93% | 95-104% |
| Note | For Research Use Only |
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