ELISA Kit for Adropin (AD)

ENHO; UNQ470; Energy Homeostasis Associated Protein

Specificity

This assay has high sensitivity and excellent specificity for detection of Adropin (AD).
No significant cross-reactivity or interference between Adropin (AD) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Adropin (AD) and the recovery rates were calculated by comparing the measured value to the expected amount of Adropin (AD) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 89-96 93
EDTA plasma(n=5) 78-98 82
heparin plasma(n=5) 79-98 83

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Adropin (AD) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Adropin (AD) 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 Adropin (AD) 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) 96-103% 85-92% 97-104% 88-102%
EDTA plasma(n=5) 88-104% 83-96% 84-92% 82-91%
heparin plasma(n=5) 82-90% 82-101% 87-98% 94-101%

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 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

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Magazine Citations
Pregnancy Hypertens. Alteration of serum adropin level in preeclampsia. pubmed:28501281
Frontiers in Molecular Neuroscience Adropin Is a Key Mediator of Hypoxia Induced Anti-Dipsogenic Effects via TRPV4-CamKK-AMPK Signaling in the Circumventricular Organs of Rats. pubmed:28473751
Preprints Afamin and Adropin in Patients with Alcohol-Induced Liver Cirrhosis manuscript:201803.0258
Probiotics and Antimicrobial Proteins  Effects of Probiotic Yogurt on Serum Omentin-1, Adropin, and Nesfatin-1 Concentrations in Overweight and Obese Participants Under Low-Calorie Diet Pubmed: 30232744
Probiotics and Antimicrobial Proteins Mitra Zarrati, Mahsa Raji Lahiji, Eisa Salehi, Bahareh Yazdani, Elham Razmpoosh, Raheleh Shokouhi Shoormasti & Farzad Shidfar Pubmed: 30232744
Pediatr Diabetes Adropin and apelin‐12 efficiently predict metabolic syndrome in obese children Pubmed: 32749012
European Journal of Therapeutics Effect of Peri-Implant Disease on Adropin Levels: A Cross-Sectional Pilot Study
Horm Mol Biol Clin Investig Effects of eight weeks exercise training on serum levels of adropin in male volleyball players 33794077
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