Overview
Product name: |
Human IL18(Interleukin 18) ELISA Kit |
Reactivity: |
Human |
Alternative Names: |
IL-18?IGIF; IL-1g; IL1F4; IL1-F4; Interferon-Gamma-Inducing Factor; Interleukin-1 Family Member 4; Iboctadekin; Interleukin-1 Gamma |
Assay Type: |
Sandwich |
Sensitivity: |
5.9 pg/mL |
Standard: |
1000 pg/mL |
Detection Range: |
15.63-1000 pg/mL |
Sample Type: |
serum, plasma, urine, 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 IL18. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human IL18. 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 IL18, 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 IL18 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 |
2.074 |
1.959 |
500.00 |
1.622 |
1.507 |
250.00 |
1.273 |
1.158 |
125.00 |
0.968 |
0.853 |
62.50 |
0.542 |
0.427 |
31.25 |
0.356 |
0.241 |
15.63 |
0.217 |
0.102 |
0.00 |
0.115 |
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 IL18 and the recovery rates were calculated by comparing the measured value to the expected amount of IL18 in samples.
Matrix |
Recovery range |
Average |
serum(n=5) |
87-95% |
91% |
EDTA plasma(n=5) |
78-92% |
85% |
Heparin plasma(n=5) |
92-106% |
99% |
Linearity
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of IL18 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% |
91-104% |
87-98% |
95-99% |
EDTA plasma(n=5) |
89-99% |
83-95% |
88-94% |
96-101% |
Heparin plasma(n=5) |
85-100% |
84-98% |
95-101% |
88-102% |