ELISA Kit for Fibronectin Type III Domain Containing Protein 5 (FNDC5)

FRCP2; Irisin; Fibronectin type III repeat-containing protein 2

Specificity

This assay has high sensitivity and excellent specificity for detection of Fibronectin Type III Domain Containing Protein 5 (FNDC5).
No significant cross-reactivity or interference between Fibronectin Type III Domain Containing Protein 5 (FNDC5) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Fibronectin Type III Domain Containing Protein 5 (FNDC5) and the recovery rates were calculated by comparing the measured value to the expected amount of Fibronectin Type III Domain Containing Protein 5 (FNDC5) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 98-105 101
EDTA plasma(n=5) 81-94 84
heparin plasma(n=5) 87-96 91

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Fibronectin Type III Domain Containing Protein 5 (FNDC5) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Fibronectin Type III Domain Containing Protein 5 (FNDC5) 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 Fibronectin Type III Domain Containing Protein 5 (FNDC5) 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) 85-104% 79-99% 84-99% 83-97%
EDTA plasma(n=5) 89-98% 80-94% 92-99% 86-101%
heparin plasma(n=5) 78-94% 80-90% 85-93% 98-105%

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
Diabetes Research and Clinical Practice Serum irisin levels in new-onset type 2 diabetes PubMed: 23369227
Journal of diabetes and its complications Lower circulating irisin is associated with type 2 diabetes mellitus Pubmed: 23619195
Horm Metab Res. Inconsistency in circulating irisin levels: what is really happening Pubmed:24459033
Int J Obes (Lond). Leptin administration activates irisin-induced myogenesis via nitric oxide-dependent mechanisms, but reduces its effect on subcutaneous fat browning in mice Pubmed:25199621
Molecular and Cellular Endocrinology Fenofibrate (PPARalpha agonist) induces beige cell formation in subcutaneous white adipose tissue from diet-induced male obese mice Pubmed:25576856
journal of diabetes Lower irisin level in patients with type 2 diabetes mellitus: A case-control study and meta-analysis PubMed: 25494632
The Journal of Maternal-Fetal & Neonatal Medicine Maternal circulating levels of irisin in intrahepatic cholestasis of pregnancy PubMed: 26689349
Hormone and Metabolic Research Lower Circulating Irisin Level in Patients with Diabetes Mellitus: A Systematic Review and Meta-Analysis Pubmed:27300472
Nature Reviews Endocrinology Physiology and role of irisin in glucose homeostasis. pubmed:28211512
Journal of Molecular and Cellular Cardiology Irisin improves perivascular adipose tissue dysfunction via regulation of the heme oxygenase-1/adiponectin axis in diet-induced obese mice pubmed:27638193
Biochemical and Biophysical Research Communications Irisin stimulates cell proliferation and invasion by targeting the PI3K/AKT pathway in human hepatocellular carcinoma pubmed:28867187
PLoS One Differential actions of PPAR-α and PPAR-β/δ on beige adipocyte formation: A study in the subcutaneous white adipose tissue of obese male mice Pubmed:29351550
International Brazilian Journal of Urology The Diagnostic Value of FNDC5/Irisin in Renal Cell Cancer. Pubmed:29522296
Redox Biology Irisin alleviates liver ischemia-reperfusion injury by inhibiting excessive mitochondrial fission, promoting mitochondrial biogenesis and decreasing oxidative … Doi: 10.1016/j.redox.2018.10.019
Antioxidants & redox signaling Low serum irisin concentration is associated with poor outcomes in patients with acute pancreatitis and irisin administration protects against experimental acute … Pubmed: 31250660
Journal of Maternal-Fetal & Neonatal Medicine Maternal serum, placental, and umbilical venous blood irisin levels in intrahepatic cholestasis of pregnancy Pubmed: 31590596
Biomed Pharmacother Identification of irisin as a therapeutic agent that inhibits oxidative stress and fibrosis in a murine model of chronic pancreatitis Pubmed: 32199226
Irisin Improves Autophagy of Aged Hepatocytes via Increasing Telomerase Activity in Liver Injury Pubmed: 31976032
Serum Irisin Predicts Posthepatectomy Complications in Patients with Hepatocellular Carcinoma Pubmed: 31976024
DOMESTIC ANIMAL ENDOCRINOLOGY The myokine Irisin: localization and effects in swine late medium and large antral ovarian follicle Pubmed: 33120167
Journal of Biomaterials and Tissue Engineering The Combination of Aerobic and Resistance Exercise Induces Weight Loss via the PGC-1α/Irisin/UCP-1 Pathway
Evaluation of a recombinant bacillus calmette-guérin vaccine expressing P39-L7/L12 of Brucella melitensis: An immunization strategy against brucellosis in BALB/c …
Eur Rev Med Pharmacol Sci Salivary Irisin: potential inflammatory biomarker in recurrent apthous stomatitis patients 33755963
Metabolites Pattern of Adiponectin, Osteocalcin, Irisin, FGF-21, and MCP-1 According to the Body Size Phenotype: Could They Be Markers of Metabolic Health in Mexican-Mestizo … 34822430
Eat Weight Disord Time-restricted feeding prevents metabolic diseases through the regulation of galanin/GALR1 expression in the hypothalamus of mice 34370270
Ir J Med Sci Eight-week exercise program improved the quality of life of Alzheimer's patients through functional, cognitive, and biochemical parameters Pubmed:35604540
Biomed Pharmacother Irisin attenuates sepsis-induced cardiac dysfunction by attenuating inflammation-induced pyroptosis through a mitochondrial ubiquitin ligase-dependent … Pubmed:35653888
Journal of King Saud University - Science Interaction of nitric oxide synthase and mitochondrial ATP-sensitive potassium channels in protective impacts of combination therapy with irisin …
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