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Manual Download| Product Name | FS-Human INS(Insulin) ELISA Kit | Species | Human |
| Uniprot ID | P01308 | Alternative Names | IDDM2, ILPR, INS, Insulin, IRDN, MODY10, proinsulin |
| Detection method | Sandwich | Sensitivity | 0.16 μIU/mL |
| Standard | 20μIU/mL | Detection Range | 0.31-20μIU/mL |
| Sample type | Serum, Plasma, Tissue homogenates, Cell lysates, Cell culture supernates and Other biological fluids;Sample Volume=50μL | ||
| Reaction time | 1.5H | Research Area | Cancer, Immunology; |
| Test principle | This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human INS. Samples (or Standards) and Horseradish Peroxidase (HRP) linked antibody specific for Human INS are added to the micro ELISA plate wells. Human INS in samples (or standards) combines with the coated antibody and HRP linked detection antibody special to Human INS. Excess conjugate and unbound sample or standard are washed from the plate. The substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450±2 nm. The OD value is proportional to the concentration of Human INS. The concentration of Human INS in the samples is then determined by comparing the OD of the samples to the standard curve. | ||
As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
| (μIU/mL) | OD | Corrected |
|---|---|---|
| 20.00 | 2.418 | 2.360 |
| 10.00 | 1.663 | 1.605 |
| 5.00 | 0.998 | 0.940 |
| 2.50 | 0.518 | 0.460 |
| 1.25 | 0.266 | 0.208 |
| 0.63 | 0.136 | 0.078 |
| 0.32 | 0.105 | 0.047 |
| 0.00 | 0.058 |
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level were tested on 3 different plates, 20 replicates in each plate, respectively.
| Intra-assay Precision | Inter-assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean(μIU/mL) | 0.61 | 4.30 | 8.18 | 0.60 | 4.39 | 8.59 |
| Standard deviation | 0.03 | 0.21 | 0.57 | 0.02 | 0.35 | 0.64 |
| C V (%) | 4.92 | 4.88 | 6.97 | 3.33 | 7.97 | 7.45 |
The recovery of spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
| Sample Type | Range(%) | Average Recovery(%) |
|---|---|---|
| Serum (n=8) | 87-95 | 91 |
| EDTA plasma (n=8) | 85-97 | 91 |
| Cell culture media (n=8) | 82-97 | 89 |
Samples were spiked with high concentrations of target proteins 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 (%) | 95-103 | 95-103 | 80-96 |
| Average (%) | 101 | 101 | 87 | |
| 1:4 | Range (%) | 80-97 | 95-103 | 88-98 |
| Average (%) | 91 | 102 | 93 | |
| 1:8 | Range (%) | 93-102 | 87-98 | 87-103 |
| Average (%) | 94 | 92 | 97 | |
| 1:16 | Range (%) | 87-103 | 85-92 | 89-97 |
| Average (%) | 94 | 88 | 95 |
