Components:
HSA Calibration Standard
Anti-HSA Microtiter Strips
Diluent
Wash Buffer Concentrate (10×)
Anti-HSA:HRP (100×)
TMB Substrate
Stop Solution
Sealing Film
This kit is based on the solid-phase enzyme-linked immunosorbent assay (ELISA) with a double-antibody sandwich technique to determine the amount of hIL-6 in biological products. A hIL-6-specific capture antibody was pre-coated into each well of microtiter strips. Both Calibration Standards and test samples were simultaneously added to the microtiter strips, and followed by incubation and washing. The biotinylated detection antibody was added to the microtiter strips to bind the epitope of hIL-6, thus forming a sandwich structure, which further reacted with streptavidin-labeled HRP (SA-HRP). TMB substrate was added into reaction, catalyzed by enzymatic hydrolysis to produce a blue colored product (maximum absorption peak at 655 nm). Finally, a stop solution was added to terminate the enzymatic reaction, resulting in a yellow colored product (maximum absorption peak at 450 nm). The absorbance values at 450 nm wavelength were positively correlated with the hIL-6 concentration in the Calibration Standards and the samples. The concentration of hIL-6 in the sample can be calculated using a dose-response curve.
| Linearity & Range | 2-512 pg/mL, R2>0.990 | ||
| Accuracy | Recovery Rate= 96.3%-109.8% | ||
| LLOQ | 2 pg/mL | ||
| ULOQ | 512 pg/mL | ||
| Detection limit | 0.7 pg/mL | ||
| Precision | 2.4%-6.7% (CV≤20%) | ||
| Specificity | No cross-reactivity with Human IL-2, Human IL-10, Human IL-12, Human IFN-γ, Human IFN-β, Human IL-6Rα, Rat IL-6, Mouse IL-6. | ||
| Robustness | Incubate at 25±5℃, CV≤ 20% | ||
| Instrument suitability not limited to MD Spectra Max ABS and Thermo Multiskan FC | |||
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