SingaporeSG
ChinaCN
MalaysiaMY
IndonesiaID
MyanmarMM_pddqnqv8.png)
Bio Reagents & Consumables
ELISA (Enzyme-linked Immunosorbent Assay)
ELK1036 Human IFNg (Interferon Gamma) ELISA Kit
Overview
| Product name: | Human IFNg(Interferon Gamma) ELISA Kit |
| Reactivity: | Human |
| Alternative Names: | IFN gamma; IFN-G; IFG; IFI; INFr; IFN; Immune Interferon |
| Assay Type: | Sandwich |
| Sensitivity: | 5.9 pg/mL |
| Standard: | 1000 pg/mL |
| Detection Range: | 15.63-1000 pg/mL |
| Sample Type: | serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids |
| Assay Length: | 3.5h |
| Research Area: | Cytokine;Infection immunity; |
| 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 Human IFNg. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human IFNg. 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 Human IFNg, 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 Human IFNg in the samples is then determined by comparing the OD of the samples to the standard curve. |
Standard curve
| Concentration (ng/mL) | OD | Corrected OD |
|---|---|---|
| 1000.00 | 2.128 | 1.998 |
| 500.00 | 1.482 | 1.352 |
| 250.00 | 0.896 | 0.766 |
| 125.00 | 0.543 | 0.413 |
| 62.50 | 0.306 | 0.176 |
| 31.25 | 0.251 | 0.121 |
| 15.63 | 0.222 | 0.092 |
| 0.00 | 0.130 | 0.000 |
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 IFNg and the recovery rates were calculated by comparing the measured value to the expected amount of IFNg in samples.
| Matrix | Recovery range | Average |
|---|---|---|
| serum(n=5) | 96-107% | 101% |
| EDTA plasma(n=5) | 87-99% | 92% |
| Heparin plasma(n=5) | 84-96% | 90% |
Linearity
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of IFNg 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) | 86-93% | 91-103% | 87-98% | 95-101% |
| EDTA plasma(n=5) | 89-99% | 83-95% | 98-104% | 96-106% |
| Heparin plasma(n=5) | 85-100% | 84-98% | 95-101% | 88-102% |
| Details | Kindly refer to the "Overview" tab. |
Author: Jianglong Li, Xiangmin Li, Hui Ma, Xujiao Ren, Genxi Hao, Huawei Zhang, Zekai Zhao, Kui Fang, Xinxin Li, Zhenxiang Rong, Shaohua Sun, Huanchun Chen, Ping Qian
Publication: Vaccine
IF: 3.641
Classical swine fever (CSF) remains one of the most important highly contagious and fatal viral disease of swine with high morbidity and mortality. CSF is caused by classical swine fever virus (CSFV), a small, enveloped RNA virus of the genus Pestivirus. The aim of this study was to construct the a novel CSFV Fc-fusion recombinant protein and evaluate the efficacy as a vaccine against CSFV. Here, we obtained a novel subunit vaccine expressing CSFV E2 recombinant fusion protein in CHO-S cells. Functional analysis revealed that CSFV Fc-fusion recombinant protein (CSFV-E2-Fc) could bind to FcγRI on antigen-presenting cells (APCs) and significantly increase IgA levels in serum and feces, inducing stronger mucosal immune response in swine. Additionally, CSFV-E2-Fc immunization enhanced CSFV-specific T cell immune response with a Th1-like pattern of cytokine secretion, remarkably stimulated the Th1-biased cellular immune response and humoral immune response. Further, the protective effects of CSFV-E2-Fc subunit vaccines were confirmed. The data suggest that CSFV E2-Fc recombinant fusion protein may be a promising candidate subunit vaccine to elicit immune response and protect against CSFV.