Multiplex Assay Kit for Fibronectin Type III Domain Containing Protein 5 (FNDC5) ,etc. by FLIA (Flow Luminescence Immunoassay) Homo sapiens (Human) Multiplex ELISA

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

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

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  • Multiplex Assay Kit for Fibronectin Type III Domain Containing Protein 5 (FNDC5) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Fibronectin Type III Domain Containing Protein 5 (FNDC5) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Fibronectin Type III Domain Containing Protein 5 (FNDC5) ,etc. by FLIA (Flow Luminescence Immunoassay) Results demonstration
  • LMN576Hu.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) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Fibronectin Type III Domain Containing Protein 5 (FNDC5) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 91-99 94
EDTA plasma(n=5) 81-98 93
heparin plasma(n=5) 98-105 101
sodium citrate plasma(n=5) 89-97 93

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Fibronectin Type III Domain Containing Protein 5 (FNDC5) ,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 Fibronectin Type III Domain Containing Protein 5 (FNDC5) ,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 Fibronectin Type III Domain Containing Protein 5 (FNDC5) ,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) 82-102% 80-91% 93-101% 96-103%
EDTA plasma(n=5) 84-97% 84-91% 91-104% 80-95%
heparin plasma(n=5) 80-89% 91-99% 92-105% 81-98%
sodium citrate plasma(n=5) 80-96% 90-98% 79-102% 96-105%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:FNDC5) 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.

Multiplex Assay Kit for Fibronectin Type III Domain Containing Protein 5 (FNDC5) ,etc. by FLIA (Flow Luminescence Immunoassay)

Test principle

Analyte-specific antibodies are pre-coated onto color-coded microparticles. Microparticles, standards, and samples are pipetted into wells and the immobilized antibodies bind the analytes of interest. After washing away any unbound substances, a biotinylated antibody cocktail specific to the analytes of interest is added to each well. Following a wash to remove any unbound biotinylated antibody, Streptavidin-Phycoerythrin conjugate (Streptavidin-PE), which binds to the biotinylated detection antibodies, is added to each well. A final wash removes unbound Streptavidin-PE and the microparticles are resuspended in buffer and read using the Luminex or Bio-Plex analyzer.The MFI developed is proportional to the concentration of analytes of interest in the sample.

Giveaways

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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