Human Interleukin 6;IL-6B-cell stimulatory factor 2 ;BSF-2;IFNB2 ELISA Kit

• For research use only.
• Cat. NO.In-Hu2192
• For the quantitative in vitro determination of Interleukin 6(IL-6) concentrations in Human serum, plasma, culture media or any biological fluid.
• Expiration date:6 months .
• Storage: 2-8℃.
• Package Size :96 wells/Kit

Principle

This ELISA kit uses Sandwich-ELISA as the method. The Microelisa stripplate provided in this kit has been pre-coated with an antibody specific to Human Interleukin 6(IL-6) . Standards or samples are added to the appropriate Microelisa stripplate wells and combined to the specific antibody. Then a Horseradish Peroxidase (HRP)- conjugated antibody specific for Human Interleukin 6(IL-6) is added to each Microelisa stripplate well and incubated. Free components are washed away. The TMB substrate solution is added to each well. Only those wells that contain Human Interleukin 6(IL-6) and HRP conjugated Human Interleukin 6(IL-6) antibody will appear blue in color and then turn yellow after the addition of the stop solution. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm. The OD value is proportional to the concentration of Human Interleukin 6(IL-6) . You can calculate the concentration of Human Interleukin 6(IL-6) in the samples by comparing the OD of the samples to the standard curve.

Materials provided with the kit

Dilution of Standards

Ten wells are set for standards in a Microelisa stripplate. In Well 1 and Well 2, 100μl Standard solution and 50μl Standard Dilution buffer are added and mixed well. In Well 3 and Well 4, 100μl solution from Well 1 and Well 2 are added respectively. Then 50μl Standard Dilution buffer are added and mixed well. 50μl solution is discarded from Well 3 and Well 4. In Well 5 and Well 6, 50μl solution from Well 3 and Well 4 are added respectively. Then 50μl Standard Dilution buffer are added and mixed well. In Well 7 and Well 8, 50μl solution from Well 5 and Well 6 are added respectively. Then 50μl Standard Dilution buffer are added and mixed well. In Well 9 and Well 10, 50μl solution from Well 7 and Well 8 are added respectively. Then 50μl Standard Dilution buffer are added and mixed well. 50μl solution is discarded from Well 9 and Well 10. After dilution, the total volume in all the wells are 50μl and the concentrations are 60 pg/ml,40pg/ml,20 pg/ml,10pg/ml and 5pg/ml, respectively.

Calculation of Results

Known concentrations of Human Interleukin 6(IL-6) Standard and its corresponding reading OD is plotted on the log scale (x-axis) and the log scale (y-axis) respectively. The concentration of Human Interleukin 6(IL-6) in sample is determined by plotting the sample’s O.D. on the Y-axis. The original concentration is calculated by multiplying the dilution factor.

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Human Interleukin 6(IL-6) were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Human Interleukin 6(IL-6) were tested on 3 different plates, 8 replicates in each plate.

CV(%) = SD/meanX100

Intra-Assay: CV<10%

Inter-Assay: CV<12%

Assay range

1pg/ml -70pg/ml

Sensitivity:

0.1 pg/ml