ELISA Kit for Fibronectin Type III Domain Containing Protein 5 (FNDC5) Mus musculus (Mouse) Sandwich ELISA

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

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  • ELISA Kit for Fibronectin Type III Domain Containing Protein 5 (FNDC5) Packages (Simulation)
  • ELISA Kit for Fibronectin Type III Domain Containing Protein 5 (FNDC5) Packages (Simulation)
  • ELISA Kit for Fibronectin Type III Domain Containing Protein 5 (FNDC5) Results demonstration
  • SEN576Mu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

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) 85-92 89
EDTA plasma(n=5) 82-103 92
heparin plasma(n=5) 92-101 97

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) 85-99% 94-101% 82-89% 80-105%
EDTA plasma(n=5) 98-105% 89-103% 82-105% 80-98%
heparin plasma(n=5) 92-99% 93-101% 97-105% 80-101%

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.

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

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Fibronectin Type III Domain Containing Protein 5 (FNDC5). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Fibronectin Type III Domain Containing Protein 5 (FNDC5). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Fibronectin Type III Domain Containing Protein 5 (FNDC5), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Fibronectin Type III Domain Containing Protein 5 (FNDC5) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Serum irisin levels in new-onset type 2 diabetesPubMed: 23369227
  • Lower circulating irisin is associated with type 2 diabetes mellitusPubmed: 23619195
  • Inconsistency in circulating irisin levels: what is really happeningPubmed:24459033
  • Leptin administration activates irisin-induced myogenesis via nitric oxide-dependent mechanisms, but reduces its effect on subcutaneous fat browning in micePubmed:25199621
  • Fenofibrate (PPARalpha agonist) induces beige cell formation in subcutaneous white adipose tissue from diet-induced male obese micePubmed:25576856
  • Lower irisin level in patients with type 2 diabetes mellitus: A case-control study and meta-analysisPubMed: 25494632
  • Maternal circulating levels of irisin in intrahepatic cholestasis of pregnancyPubMed: 26689349
  • Lower Circulating Irisin Level in Patients with Diabetes Mellitus: A Systematic Review and Meta-AnalysisPubmed:27300472
  • Physiology and role of irisin in glucose homeostasis.pubmed:28211512
  • Irisin improves perivascular adipose tissue dysfunction via regulation of the heme oxygenase-1/adiponectin axis in diet-induced obese micepubmed:27638193
  • Irisin stimulates cell proliferation and invasion by targeting the PI3K/AKT pathway in human hepatocellular carcinoma pubmed:28867187
  • Differential actions of PPAR-α and PPAR-β/δ on beige adipocyte formation: A study in the subcutaneous white adipose tissue of obese male micePubmed:29351550
  • The Diagnostic Value of FNDC5/Irisin in Renal Cell Cancer.Pubmed:29522296
  • 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
  • Low serum irisin concentration is associated with poor outcomes in patients with acute pancreatitis and irisin administration protects against experimental acute …Pubmed: 31250660
  • Maternal serum, placental, and umbilical venous blood irisin levels in intrahepatic cholestasis of pregnancyPubmed: 31590596
  • Identification of irisin as a therapeutic agent that inhibits oxidative stress and fibrosis in a murine model of chronic pancreatitisPubmed: 32199226
  • Irisin Improves Autophagy of Aged Hepatocytes via Increasing Telomerase Activity in Liver InjuryPubmed: 31976032
  • Serum Irisin Predicts Posthepatectomy Complications in Patients with Hepatocellular CarcinomaPubmed: 31976024
  • The myokine Irisin: localization and effects in swine late medium and large antral ovarian folliclePubmed: 33120167
  • 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 …
  • Salivary Irisin: potential inflammatory biomarker in recurrent apthous stomatitis patients33755963
  • 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
  • Time-restricted feeding prevents metabolic diseases through the regulation of galanin/GALR1 expression in the hypothalamus of mice34370270
  • Eight-week exercise program improved the quality of life of Alzheimer's patients through functional, cognitive, and biochemical parametersPubmed:35604540
  • Irisin attenuates sepsis-induced cardiac dysfunction by attenuating inflammation-induced pyroptosis through a mitochondrial ubiquitin ligase-dependent …Pubmed:35653888
  • Interaction of nitric oxide synthase and mitochondrial ATP-sensitive potassium channels in protective impacts of combination therapy with irisin …

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