Products specifications
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Species
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Human
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Format
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96T
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Sample Type
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Serum, plasma, tissue homogenates and other biological fluids.
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Assay Length
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4.5 h
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Detection Range
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0.156-10ng/mL. The standard curve concentrations used for the ELISA’s were 10ng/mL, 5ng/mL, 2.5ng/mL, 1.25ng/mL, 0.625ng/mL, 0.313ng/mL, 0.156ng/mL
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Sensitivity
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0.055ng/mL
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Specificity
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This assay has high sensitivity and excellent specificity for detection of D2-Hydroxyglutarate Dehydrogenase (D2HGDH).
No significant cross-reactivity or interference between D2-Hydroxyglutarate Dehydrogenase (D2HGDH) and analogues was observed.
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Intra-Assay Precision
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<10%
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Inter-Assay Precision
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<12%
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Storage Temperature
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-20 °C
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Shipping Conditions
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2-8 °C
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Handling Instruction
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The activity loss rate (stability) of this kit is less than 5% within the expiration date under appropriate storage conditions. To minimize extra influence on the stability, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. When not in use, kit components should be refrigerated. All reagents should be brought to room temperature before use. It is recommended that all standards, controls, and samples be run in duplicate. Do not mix or interchange different reagent lots from various kit lots.
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Test Principle
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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 D2-Hydroxyglutarate Dehydrogenase (D2HGDH). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to D2-Hydroxyglutarate Dehydrogenase (D2HGDH). 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 D2-Hydroxyglutarate Dehydrogenase (D2HGDH), 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 D2-Hydroxyglutarate Dehydrogenase (D2HGDH) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
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Notes
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All kits are custom made on-demand. Please allow 10-15 working days for production and 2-3 days for shipment.
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Legal Information
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Marketed under license from USCN Life Science Inc.
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| Reagents | Quantity |
| Pre-coated, ready to use 96-well strip plate | 1 |
| Standard | 2 |
| Detection Reagent A | 1×120µL |
| Detection Reagent B | 1×120µL |
| TMB Substrate | 1×9mL |
| Wash Buffer (30 × concentrate) | 1×20mL |
| Plate sealer for 96 wells | 4 |
| Standard Diluent | 1×20mL |
| Assay Diluent A | 1×12mL |
| Assay Diluent B | 1×12mL |
| Stop Solution | 1×6mL |
| Instruction manual | 1 |
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of D2-Hydroxyglutarate Dehydrogenase (D2HGDH) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
| Sample | 1:2 | 1:4 | 1:8 | 1:16 |
| serum(n=5) | 80-103% | 85-105% | 78-102% | 84-98% |
| EDTA plasma(n=5) | 79-92% | 79-104% | 84-98% | 89-101% |
| heparin plasma(n=5) | 93-101% | 89-99% | 79-94% | 92-101% |
Matrices listed below were spiked with certain level of recombinant D2-Hydroxyglutarate Dehydrogenase (D2HGDH) and the recovery rates were calculated by comparing the measured value to the expected amount of D2-Hydroxyglutarate Dehydrogenase (D2HGDH) in samples.
| Matrix | Recovery range (%) | Average(%) |
| serum(n=5) | 96-105 | 101 |
| EDTA plasma(n=5) | 89-96 | 93 |
| heparin plasma(n=5) | 79-105 | 102 |