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ELK Biotechnology
SKU:ELK7578
Mouse UBE2I(Ubiquitin Conjugating Enzyme E2I) ELISA Kit
Mouse UBE2I(Ubiquitin Conjugating Enzyme E2I) ELISA Kit
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Mouse UBE2I(Ubiquitin Conjugating Enzyme E2I) ELISA Kit
Overview
| Product name: | Mouse UBE2I(Ubiquitin Conjugating Enzyme E2I) ELISA Kit |
| Reactivity: | Mouse |
| Alternative Names: | P18; UBC9; UBCE9; SUMO-protein ligase; Ubiquitin carrier protein 9; Ubiquitin carrier protein I; Ubiquitin-protein ligase I; SUMO-conjugating enzyme UBC9 |
| Assay Type: | Sandwich |
| Sensitivity: | 0.125 ng/mL |
| Standard: | 20 ng/mL |
| Range: | 0.32-20 ng/mL |
| Sample Type: | Tissue homogenates, cell lysates and other biological fluids |
| Assay Length: | 3.5h |
| Research Area: | Enzyme & Kinase; |
| Test principle: | The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Mouse UBE2I. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Mouse UBE2I. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Mouse UBE2I, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Mouse UBE2I in the samples is then determined by comparing the OD of the samples to the standard curve. |
Standard curve
| Concentration (ng/mL) | OD | Corrected OD |
|---|---|---|
| 20.00 | 2.155 | 2.072 |
| 10.00 | 1.527 | 1.444 |
| 5.00 | 1.027 | 0.944 |
| 2.50 | 0.866 | 0.783 |
| 1.25 | 0.573 | 0.490 |
| 0.63 | 0.364 | 0.281 |
| 0.32 | 0.227 | 0.144 |
| 0.00 | 0.083 | 0.000 |

Precision
Intra-assay Precision (Precision within an assay):CV%<8%
Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-assay Precision (Precision between assays):CV%<10%
Three samples of known concentration were tested in forty separate assays to assess inter-assay precision.
Recovery
Matrices listed below were spiked with certain level of recombinant UBE2I and the recovery rates were calculated by comparing the measured value to the expected amount of UBE2I in samples.
| Matrix | Recovery range | Average |
|---|---|---|
| serum(n=5) | 84-95% | 88% |
| EDTA plasma(n=5) | 82-97% | 92% |
| Heparin plasma(n=5) | 87-99% | 94% |
Linearity
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of UBE2I and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
| Matrix | 1:2 | 1:4 | 1:8 | 1:16 |
|---|---|---|---|---|
| serum(n=5) | 89-99% | 83-95% | 98-104% | 96-106% |
| EDTA plasma(n=5) | 84-94% | 85-103% | 94-105% | 92-101% |
| Heparin plasma(n=5) | 88-104% | 84-98% | 95-105% | 88-102% |
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