Mini Samples ELISA Kit for Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF)

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Mini Samples Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF).
No significant cross-reactivity or interference between Mini Samples Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF) and analogues was observed.

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 83-96 93
EDTA plasma(n=5) 93-101 97
heparin plasma(n=5) 86-103 91

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Mini Samples Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Mini Samples Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF) 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 Mini Samples Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF) 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) 94-103% 83-96% 79-94% 93-104%
EDTA plasma(n=5) 83-90% 80-94% 78-95% 99-105%
heparin plasma(n=5) 89-96% 84-98% 78-96% 95-103%

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
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×60µL Assay Diluent A 1×6mL
Detection Reagent B 1×60µL Assay Diluent B 1×6mL
TMB Substrate 1×4.5mL Stop Solution 1×3mL
Wash Buffer (30 × concentrate) 1×10mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 25µL standard or sample to each well. Incubate 1 hour at 37°C;
3. Aspirate and add 25µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 25µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 25µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 20µL Stop Solution. Read at 450nm immediately.

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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
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