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SKU:LMC300Hu
Magnetic Luminex Assay Kit for Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF) ,etc.
Magnetic Luminex Assay Kit for Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF) ,etc.
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Product No.
LMC300Hu
Organism Species
Homo sapiens (Human).
Sample Type
Serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
Test Method
Double-antibody Sandwich
Assay Length
3.5h
Detection Range
0.01-10ng/mL
Sensitivity
The minimum detectable dose of this kit is typically less than 0.003 ng/mL.
UOM
1Plex 2Plex 3Plex 4Plex 5Plex 6Plex 7Plex 8Plex.
Specificity
This assay has high sensitivity and excellent specificity for detection of Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF) ,etc..
No significant cross-reactivity or interference between Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF) ,etc. and analogues was observed.
Recovery
Matrices listed below were spiked with certain level of recombinant Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF) ,etc. and the recovery rates were calculated by comparing the measured value to the expected amount of Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF) ,etc. in samples.
| Matrix | Recovery range (%) | Average(%) |
| serum(n=5) | 88-101 | 95 |
| EDTA plasma(n=5) | 81-90 | 85 |
| heparin plasma(n=5) | 82-103 | 92 |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF) ,etc. were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF) ,etc. 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 Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF) ,etc. 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) | 92-99% | 97-104% | 82-96% | 96-105% |
| EDTA plasma(n=5) | 84-93% | 82-96% | 80-90% | 96-104% |
| heparin plasma(n=5) | 97-105% | 82-92% | 80-94% | 94-103% |
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#:MANF) | 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 100μL standard or sample to each well,
add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.
| Magazine | Citations |
| PLoS One. | Molecular Effects of Doxycycline Treatment on Pterygium as Revealed by Massive Transcriptome Sequencing PubMed: PMC3378547 |
| Endocr J. | Circulating mesencephalic astrocyte-derived neurotrophic factor is increased in newly diagnosed prediabetic and diabetic patients, and is associated with insulin resistance. pubmed:28216543 |
| Journal of Steroid Biochemistry and Molecular Biology | Depressive-like effect of prenatal exposure to DDT involves global DNA hypomethylation and impairment of GPER1/ESR1 protein levels but not ESR2 and AHR/ARNT signaling pubmed:28263910 |
| Endocrine Journal | Circulating mesencephalic astrocyte-derived neurotrophic factor is increased in newly diagnosed prediabetic and diabetic patients, and is associated with insulin resistance 10.1507:endocrj.EJ16-0472 |
| Nature Metabolism | MANF regulates metabolic and immune homeostasis in ageing and protects against liver damage |
| J Exp Med | Feeding-induced hepatokine, Manf, ameliorates diet-induced obesity by promoting adipose browning via p38 MAPK pathway 33856409 |
| J Inflamm Res | The Potential Protective Effect of Mesencephalic Astrocyte-Derived Neurotrophic Factor on Post-Operative Delirium via Inhibiting Inflammation and Microglia … 34234505 |
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