Description
| Product name: | Human NFKB-p65(Nuclear Factor Kappa B p65) ELISA Kit |
| Reactivity: | Human |
| Alternative Names: | NFKB3, p65, RelA |
| Assay Type: | Sandwich |
| Sensitivity: | 0.10 ng/mL |
| Standard: | 10 ng/mL |
| Range: | 0.16-10 ng/mL |
| Sample Type: | serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids |
| Assay Length: | 3.5h |
| Research Area: | Signal transduction; |
| 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 Nuclear Factor Kappa B p65(NFKB-p65). Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Nuclear Factor Kappa B p65(NFKB-p65). 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 Nuclear Factor Kappa B p65(NFKB-p65), 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 Nuclear Factor Kappa B p65(NFKB-p65) 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 NFKB-p65 and the recovery rates were calculated by comparing the measured value to the expected amount of NFKB-p65 in samples.
| Matrix | Recovery range | Average |
|---|---|---|
| serum(n=5) | 96-107% | 101% |
| EDTA plasma(n=5) | 79-93% | 86% |
| Heparin plasma(n=5) | 80-97% | 88% |
Linearity
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of NFKB-p65 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) | 89-98% | 95-103% | 99-106% | 78-92% |
| EDTA plasma(n=5) | 82-96% | 87-103% | 91-97% | 81-93% |
| Heparin plasma(n=5) | 78-91% | 87-98% | 92-101% | 96-105% |
| Note | For Research Use Only |
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