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Multiplex Assay Kit for Bcl2/Adenovirus E1B 19kDa Interacting Protein 3 (BNIP3) ,etc. by FLIA (Flow Luminescence Immunoassay)
NIP3
(Note: Up to 8-plex in one testing reaction)
- Product No.LMJ545Ra
- Organism SpeciesRattus norvegicus (Rat) Same name, Different species.
- Sample TypeTissue homogenates, cell lysates and other biological fluids.
- Test MethodDouble-antibody Sandwich
- Assay Length4h
- Detection Range0.04-40ng/mL
- SensitivityThe minimum detectable dose of this kit is typically less than 0.013 ng/mL.
- DownloadInstruction Manual
- UOM 8Plex 7Plex 6Plex 5Plex 4Plex 3Plex 2Plex1Plex
- FOB
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Result
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Specificity
This assay has high sensitivity and excellent specificity for detection of Bcl2/Adenovirus E1B 19kDa Interacting Protein 3 (BNIP3).
No significant cross-reactivity or interference between Bcl2/Adenovirus E1B 19kDa Interacting Protein 3 (BNIP3) and analogues was observed.
Recovery
Matrices listed below were spiked with certain level of recombinant Bcl2/Adenovirus E1B 19kDa Interacting Protein 3 (BNIP3) and the recovery rates were calculated by comparing the measured value to the expected amount of Bcl2/Adenovirus E1B 19kDa Interacting Protein 3 (BNIP3) in samples.
| Matrix | Recovery range (%) | Average(%) |
| serum(n=5) | 89-97 | 93 |
| EDTA plasma(n=5) | 92-99 | 96 |
| heparin plasma(n=5) | 82-95 | 90 |
| sodium citrate plasma(n=5) | 95-105 | 99 |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Bcl2/Adenovirus E1B 19kDa Interacting Protein 3 (BNIP3) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Bcl2/Adenovirus E1B 19kDa Interacting Protein 3 (BNIP3) 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 Bcl2/Adenovirus E1B 19kDa Interacting Protein 3 (BNIP3) 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) | 98-105% | 92-101% | 85-99% | 88-102% |
| EDTA plasma(n=5) | 95-103% | 79-91% | 78-93% | 97-105% |
| heparin plasma(n=5) | 93-101% | 79-98% | 93-105% | 96-104% |
| sodium citrate plasma(n=5) | 79-98% | 85-92% | 94-104% | 80-97% |
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 |
| 96-well plate | 1 | Plate sealer for 96 wells | 4 |
| Pre-Mixed Standard | 2 | Standard Diluent | 1×20mL |
| Pre-Mixed Magnetic beads (22#:BNIP3) | 1 | Analysis buffer | 1×20mL |
| Pre-Mixed Detection Reagent A | 1×120μL | Assay Diluent A | 1×12mL |
| Detection Reagent B (PE-SA) | 1×120μL | Assay Diluent B | 1×12mL |
| Sheath Fluid | 1×10mL | Wash Buffer (30 × concentrate) | 1×20mL |
| Instruction manual | 1 |
Assay procedure summary
1. Add 200μL analysis buffer solution to each well of the plate for pre-wetting;
2. Preparation of standards, reagents and samples before the experiment;
3. Add 100μL standard or sample to each well, add 10μL magnetic beads, and incubate 120 minutes protect from light at 25°C on shaker;
4. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60 minutes protect from light at 25°C on shaker;
5. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent B, and incubate 30 minutes protect from light at 25°C on shaker;
6. Wash plate once on magnetic frame;
7. Add 100μL sheath fluid, swirl for 10 minutes, read on the machine.
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