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Multiplex Assay Kit for Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF) ,etc. by FLIA (Flow Luminescence Immunoassay)

ARMET; ARP; Arginine Rich Mutated In Early Stage Tumors; Arginine-rich protein

(Note: Up to 8-plex in one testing reaction)

Specificity

This assay has high sensitivity and excellent specificity for detection of Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 84-102 96
EDTA plasma(n=5) 90-97 93
heparin plasma(n=5) 80-99 90

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF) ,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 Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF) ,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 Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF) ,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) 84-92% 85-104% 98-105% 79-90%
EDTA plasma(n=5) 80-95% 94-102% 96-105% 82-99%
heparin plasma(n=5) 96-103% 94-103% 89-97% 92-99%

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.

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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
Catalog No. Related products for research use of Rattus norvegicus (Rat) Organism species Applications (RESEARCH USE ONLY!)
SEC300Ra ELISA Kit for Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF) Enzyme-linked immunosorbent assay for Antigen Detection.
LMC300Ra Multiplex Assay Kit for Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF) ,etc. by FLIA (Flow Luminescence Immunoassay) FLIA Kit for Antigen Detection.