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| Assay Type | Sandwich-ELISA |
| Analyte | IL-6 |
| Format | 96T(8×12 strips) |
| Reactivity | Human |
| Regulatory Status | RUO |
| Sensitivity | <2.34pg/mL |
| Standard Curve Range | 2.34 pg/mL-150 pg/mL |
| Assay Time | 2 hr 50 min |
| Suitable Sample Type | For the quantitative determination of human IL-6 in Cell Culture Supernatants, Plasma, Serum. |
| Sample volume | 100 uL |
| ID | Components | Size |
| CRS005-C01 | Pre-coated Anti-IL-6 Antibody Microplate | 1 plate(8×12 strips) |
| CRS005-C02 | Human IL-6 Standard | 20 μg |
| CRS005-C03 | Biotin-Anti-IL-6 Antibody | 50 μL |
| CRS005-C04 | Streptavidin-HRP | 50 μL |
| CRS005-C05 | 10xWashing Buffer | 50 mL |
| CRS005-C06 | 2xDilution Buffer | 50 mL |
| CRS005-C07 | Substrate Solution | 12 mL |
| CRS005-C08 | Stop Solution | 7 mL |
Your experiment will include 6 simple steps:
a) Bring all reagents to room temperature(20℃-25℃) before use.
b) Add your sample to the plate and take the Human IL-6 as standard. The samples and standard are diluted by Dilution Buffer.
c) Wash the plate and add the Biotin-Anti-IL-6 Antibody diluted by Dilution Buffer to the plate.
d) Wash the plate and add the Streptavidin-HRP diluted by Dilution Buffer to the plate.
e) Wash the plate and add TMB.
f) Stop the substrate reaction by adding diluted acid. Absorbance (OD) is calculated by the absorbance at 450 nm minus the absorbance at 630 nm to remove background disturbance before statistical analysis. The OD Value reflects the amount of bound protein.
For each experiment, a standard curve needs to be set for each micro-plate, and the specific OD value may vary depending on different laboratories, testers, or equipments. The following example data is for reference only.
To assess the linearity of the assay, samples spiked with high concentrations of human IL-6 were serially diluted with calibrator diluent to produce samples with values within the dynamic range of the assay.
Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision, Intra-Assay Precision CV＜10%.
Three samples of known concentration were tested in three separate assays to assess inter-assay precision, Inter-Assay Precision CV<10%.
Three parts of blank serum were added with different concentrations of human IL-6, and the serum without human IL-6 was used as background to calculate the recovery rate. The range of the recovery rate is 90.6-114.8%, and the average recovery is 101.2%.
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