This assay has high sensitivity and excellent specificity for detection of High Sensitive Interleukin 35 (IL35).No significant cross-reactivity or interference between High Sensitive Interleukin 35 (IL35) and analogues was observed.

Matrices listed below were spiked with certain level of recombinant High Sensitive Interleukin 35 (IL35) and the recovery rates were calculated by comparing the measured value to the expected amount of High Sensitive Interleukin 35 (IL35) in samples.

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level High Sensitive Interleukin 35 (IL35) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level High Sensitive Interleukin 35 (IL35) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100 Intra-Assay: CV<10%>Inter-Assay: CV<12%>

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of High Sensitive Interleukin 35 (IL35) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

The microplate provided in this kit has been pre-coated with a monoclonal antibody specific to IL12A. Standards or samples are then added to the appropriate microplate wells with a biotin-conjugated monoclonal antibody specific to IL27B. 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 IL35, 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 IL35 in the samples is then determined by comparing the O.D. of the samples to the standard curve.

ELISA / CLIA Experiment Service

Single-component Reagents of Assay Kit