Code | CSB-E08464h |
Size | $496 |
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Trial Size |
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Intra-assay Precision (Precision within an assay): CV%<8% | ||||||
Three samples of known concentration were tested twenty times on one plate to assess. | ||||||
Inter-assay Precision (Precision between assays): CV%<10% | ||||||
Three samples of known concentration were tested in twenty assays to assess. |
To assess the linearity of the assay, samples were spiked with high concentrations of human IL-27 in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay. | ||||||
| Sample | Serum(n=4) | ||||
1:1 | Average % | 94 | ||||
Range % | 90-98 | |||||
1:2 | Average % | 89 | ||||
Range % | 84-95 | |||||
1:4 | Average % | 92 | ||||
Range % | 87-96 | |||||
1:8 | Average % | 97 | ||||
Range % | 92-104 |
The recovery of human IL-27 spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section. | ||||||
Sample Type | Average % Recovery | Range | ||||
Serum (n=5) | 88 | 84-93 | ||||
EDTA plasma (n=4) | 90 | 88-93 |
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed. | |||||||
ng/ml | OD1 | OD2 | Average | Corrected | |||
10 | 2.815 | 2.869 | 2.842 | 2.717 | |||
5 | 2.436 | 2.647 | 2.542 | 2.417 | |||
2.5 | 1.954 | 1.968 | 1.961 | 1.836 | |||
1.25 | 1.312 | 1.336 | 1.324 | 1.199 | |||
0.625 | 0.764 | 0.704 | 0.734 | 0.609 | |||
0.312 | 0.468 | 0.442 | 0.455 | 0.330 | |||
0.156 | 0.297 | 0.274 | 0.286 | 0.161 | |||
0 | 0.124 | 0.126 | 0.125 |
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This human IL-27 ELISA Kit is suitable for qualitatively determining human concentrations in multiple biological fluids, including human serum, plasma, and tissue homogenates in vitro.IL27 is a heterodimeric cytokine that plays a role both in innate and adaptive immunity. IL27 engages the IL27 receptor and then recruits several Jak family kinases, activating the JAK-STAT pathway, thus mediating many cellular processes, including cellular differentiation, proliferation, and immunologic responses. Studies have shown that IL27 regulates T cell responses that prevent immune hyperactivity, suggesting a possible therapeutic for chronic inflammatory conditions with excessive T cell activation, such as MS and rheumatoid arthritis. Contrastly, IL27 exerts pro-inflammatory functions by promoting the clonal expansion of naive CD4+ T-cells and the survival of Treg and CD8+ T cells and stimulating tumor-specific cytotoxic T cell responses.
This kit uses the quantitative sandwich-based enzyme immunoassay technique to measure the amount of human IL-27 in the sample. Standards and samples are respectively added to the microplate wells pre-coated with an anti-human IL-27 antibody. Biotin-labeled IL-27 antibody, HRP-avidin, and TMB substrate are pipped into the microplate in turn. The capture antibody pre-coated on the plate captures the IL-27 in the human samples. IL-27 binds to the biotinylated anti-IL-27 human monoclonal antibody. And the biotin on the biotinylated anti-IL-27 human monoclonal antibody binds to the avidin on the enzyme label, forming immune complexes. The color renders blue after the addition of the TMB substrate. The addition of the stop solution into the wells immediately turns the blue into yellow. The concentration of IL-27 in the samples is directly proportional to OD (450nm). Each manufactured lot of this ELISA kit was quality tested for criteria such as sensitivity, specificity, precision, linearity, and lot-to-lot consistency.
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