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| Detection Range: | 3.13 - 200 ng/mL |
| Sensitivity: | 1.88 ng/mL |
| Species: | Rat |
| Synonym: | IDDM2, ILPR, IRDN, MODY10 |
| Background: | This ELISA kit can be applied to the in vitro quantitative determination of Rat INS concentrations in serum, plasma and other biological fluids. This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Rat INS. Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Rat INS and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Rat INS, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Rat INS. The concentration of Rat INS in samples can be calculated by comparing the OD of the samples to the standard curve. |
| Sample Volume Required Per Well: | 100 μL |
| Sample Type: | Serum, plasma and other biological fluids |
Typical data
For convenience in result calculation, absorbance can be used as ordinate and standard concentrations as abscissa. The standard curve provided in the manual is only for reference, and experimenters should draw the standard curve based on data of themselves.
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high leve Rat INS were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Rat INS were tested on 3 different plates, 20 replicates in each plate.
| Intra-assay Precision | Inter-assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (ng/mL) | 10.40 | 24.10 | 80.40 | 10.90 | 22.10 | 74.30 |
| Standard deviation | 0.60 | 1.20 | 3.10 | 0.70 | 1.00 | 2.90 |
| C V (%) | 5.77 | 4.98 | 3.86 | 6.42 | 4.52 | 3.90 |
Recovery
The recovery of Rat INS spiked to three different levels in samples throughout the range of the assay in various matrices was evaluated.
| Sample Type | Range (%) | Average Recovery (%) |
|---|---|---|
| Serum (n=5) | 89-103 | 95 |
| EDTA plasma (n=5) | 88-104 | 95 |
| Cell culture media (n=5) | 92-107 | 100 |
Linearity
Samples were spiked with high concentrations of Rat INS and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
|---|---|---|---|---|
| 1:2 | Range (%) | 85-98 | 94-108 | 94-108 |
| Average (%) | 91 | 100 | 100 | |
| 1:4 | Range (%) | 100-114 | 81-91 | 96-109 |
| Average (%) | 106 | 86 | 102 | |
| 1:8 | Range (%) | 96-108 | 86-97 | 95-108 |
| Average (%) | 102 | 91 | 102 | |
| 1:16 | Range (%) | 94-108 | 82-93 | 97-108 |
| Average (%) | 101 | 88 | 103 |
Kit components & Storage
The unopened kit can be stored at 4℃ for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions since the kit is received.
| Item | Specifications | Storage |
|---|---|---|
| Micro ELISA Plate(Dismountable) | 8 wells ×12 strips | -20℃, 6 months |
| Reference Standard | 2 vials | |
INS
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