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
| Materials provided with the kit | 48 determinations | 96 determinations | Storage |
| 1 | User manual | 1 | 1 | R.T. |
| 2 | Closure plate membrane | 2 | 2 | R.T. |
| 3 | Sealed bags | 1 | 1 | R.T. |
| 4 | Microelisa stripplate | 1 | 1 | 2-8℃ |
| 5 | Standard:90pg/ml | 0.5ml×1 bottle | 0.5ml×1 bottle | 2-8℃ |
| 6 | Standard diluent | 1.5ml×1 bottle | 1.5ml×1 bottle | 2-8℃ |
| 7 | HRP-Conjugate reagent | 3ml×1 bottle | 6ml×1 bottle | 2-8℃ |
| 8 | Sample diluent | 3ml×1 bottle | 6ml×1 bottle | 2-8℃ |
| 9 | Chromogen Solution A | 3ml×1 bottle | 6ml×1 bottle | 2-8℃ |
| 10 | Chromogen Solution B | 3ml×1 bottle | 6ml×1 bottle | 2-8℃ |
| 11 | Stop Solution | 3ml×1 bottle | 6ml×1 bottle | 2-8℃ |
| 12 | Wash solution | 20ml (20X)×1bottle | 20ml (30X)×1bottle | 2-8℃ |
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
