OneStep ELISA Kit for Diazepam Binding Inhibitor (DBI) 
ACBP; ACBD1; CCK-RP; EP; Endozepine; GABA Receptor Modulator; Acyl-Coenzyme A Binding Protein
Overview
Properties
- Product No.OED696Hu
- Organism SpeciesHomo sapiens (Human) Same name, Different species.
- ApplicationsEnzyme-linked immunosorbent assay for Antigen Detection.
Research use only - DownloadInstruction Manual
- CategoryMetabolic pathway
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Packages (Simulation)
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Packages (Simulation)
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ISO9001: 2008, ISO13485: 2003 Registered
Recovery
Matrices listed below were spiked with certain level of recombinant Diazepam Binding Inhibitor (DBI) and the recovery rates were calculated by comparing the measured value to the expected amount of Diazepam Binding Inhibitor (DBI) in samples.
| Matrix | Recovery range (%) | Average(%) |
| serum(n=5) | 92-102 | 96 |
| EDTA plasma(n=5) | 79-94 | 88 |
| heparin plasma(n=5) | 82-92 | 88 |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Diazepam Binding Inhibitor (DBI) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Diazepam Binding Inhibitor (DBI) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%
Linearity
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Diazepam Binding Inhibitor (DBI) 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) | 80-91% | 80-94% | 87-103% | 89-103% |
| EDTA plasma(n=5) | 84-105% | 94-103% | 79-99% | 78-97% |
| heparin plasma(n=5) | 80-91% | 78-93% | 89-104% | 81-101% |
Stability
The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.
Reagents and materials provided
| Reagents | Quantity | Reagents | Quantity |
Assay procedure summary
1. Prepare all reagents, samples and standards;
2. Add 50µL standard or sample to each well, then add 50µL prepared Detection Reagent A to each well. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
5. Add 50µL Stop Solution. Read at 450nm immediately.
Test principle
The microplate provided in this kit has been pre-coated with an antibody specific to DBI. Standards or samples are then added to the appropriate microplate wells with a HRP-conjugated antibody specific to DBI. After TMB substrate solution is added, only those wells that contain DBI and HRP-conjugated antibody 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 DBI in the samples is then determined by comparing the O.D. of the samples to the standard curve.
