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Multiplex Assay Kit for Mitochondrial Open Reading Frame Of The 12S rRNA-c (MOTS-c) ,etc. by FLIA (Flow Luminescence Immunoassay)
(Note: Up to 8-plex in one testing reaction)
- Product No.LMX132Hu
- Organism SpeciesHomo sapiens (Human) Same name, Different species.
- Sample TypeSerum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
- Test MethodCompetitive Inhibition
- Assay Length1.5h
- Detection Range0.2-200ng/mL
- SensitivityThe minimum detectable dose of this kit is typically less than 0.067 ng/mL.
- DownloadInstruction Manual
- UOM 8Plex 7Plex 6Plex 5Plex 4Plex 3Plex 2Plex1Plex
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Result
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Specificity
This assay has high sensitivity and excellent specificity for detection of Mitochondrial Open Reading Frame Of The 12S rRNA-c (MOTS-c) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Mitochondrial Open Reading Frame Of The 12S rRNA-c (MOTS-c) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.
Recovery
Matrices listed below were spiked with certain level of recombinant Mitochondrial Open Reading Frame Of The 12S rRNA-c (MOTS-c) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Mitochondrial Open Reading Frame Of The 12S rRNA-c (MOTS-c) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.
| Matrix | Recovery range (%) | Average(%) |
| serum(n=5) | 86-101 | 89 |
| EDTA plasma(n=5) | 78-102 | 81 |
| heparin plasma(n=5) | 79-98 | 83 |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Mitochondrial Open Reading Frame Of The 12S rRNA-c (MOTS-c) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Mitochondrial Open Reading Frame Of The 12S rRNA-c (MOTS-c) ,etc. by FLIA (Flow Luminescence Immunoassay) 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 Mitochondrial Open Reading Frame Of The 12S rRNA-c (MOTS-c) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) | 81-97% | 78-93% | 78-94% | 87-101% |
| EDTA plasma(n=5) | 93-105% | 87-102% | 87-94% | 78-103% |
| heparin plasma(n=5) | 96-105% | 94-105% | 90-99% | 86-98% |
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#:MOTS-c) | 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. Preparation of standards, reagents and samples before the experiment;
2. Add 50μL standard or sample to each well,
add 10μL magnetic beads,and 50μL Detection Reagent A,incubate 60min at 37°C on shaker;
3. Wash plate on magnetic frame for three times;
4. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
5. Wash plate on magnetic frame for three times;
6. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.
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| Magazine | Citations |
| Pediatric Diabetes | Circulating MOTS‐c levels are decreased in obese male children and adolescents and associated with insulin resistance Pubmed:29691953 |
| Rapid Commun Mass Spectrom | Development of a mass spectrometry‐based detection method for the mitochondrion‐derived peptide MOTS‐c in plasma samples for doping control purposes Pubmed: 30394592 |
| DIABETOLOGIA | Adiponectin treatment improves insulin resistance in mice by regulating the expression of the mitochondrial-derived peptide MOTS-c and its response to exercise via … Pubmed: 32880686 |
| Induction of hyperandrogenism and insulin resistance differentially modulates ferroptosis in uterine and placental tissues of pregnant rats | |
| FASEB JOURNAL | Neural cell©\derived plasma exosome protein abnormalities implicate mitochondrial impairment in first episodes of psychosis 33454965 |
| Biochim Biophys Acta Mol Basis Dis | MOTS-c interacts synergistically with exercise intervention to regulate PGC-1¦Á expression, attenuate insulin resistance and enhance glucose metabolism in mice via?¡ 33722744 |
| researchsquare | Evolution of Mitochondrial-Derived Peptides, Humanin and MOTSc and Changes in Insulin Sensitivity During Early Gestation |
| Mol Psychiatry | Abnormal levels of mitochondrial proteins in plasma neuronal extracellular vesicles in major depressive disorder 34471251 |
| Pharmacol Res | The Mitochondrial-Derived Peptide MOTS-c Relieves Hyperglycemia and Insulin Resistance in Gestational Diabetes Mellitus 34798268 |
| Ann Neurol | SARS‐CoV‐2 and mitochondrial proteins in neural‐derived exosomes of COVID‐19 Pubmed:35285072 |
| J Clin Med | Evolution of Mitochondrially Derived Peptides Humanin and MOTSc, and Changes in Insulin Sensitivity during Early Gestation in Women with and without Gestational … Pubmed:35683389 |