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ELISA Kit for Fibronectin (FN) Homo sapiens (Human) Sandwich ELISA

FN1; CIG; FINC; LETS; MSF; GFND2; Anastellin; Migration-Stimulating Factor; Cold-Insoluble Globulin; Large, External, Transformation-Sensitive Protein

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  • ELISA Kit for Fibronectin (FN) Packages (Simulation)
  • ELISA Kit for Fibronectin (FN) Packages (Simulation)
  • ELISA Kit for Fibronectin (FN) Results demonstration
  • SEA037Hu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 80-105 91
EDTA plasma(n=5) 78-101 92
heparin plasma(n=5) 78-98 84

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Fibronectin (FN) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Fibronectin (FN) 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 (FN) 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) 93-101% 98-105% 93-101% 89-99%
EDTA plasma(n=5) 95-103% 92-101% 83-93% 96-104%
heparin plasma(n=5) 85-101% 79-103% 95-104% 78-101%

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 (FN)

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 (FN). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Fibronectin (FN). 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 (FN), 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 (FN) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Advanced oxidation protein products induce mesangial cell perturbation through PKC-dependent activation of NADPH oxidasePubMed: 19019916
  • Protein synthesis and secretion in human mesenchymal cells derived from bone marrow, adipose tissue and Wharton's jellyPubmed: 24739658
  • Identification of compounds from the water soluble extract of Cinnamomum cassia barks and their inhibitory effects against high-glucose-induced mesangial cells.Pubmed: 24013407
  • Bioactive compounds from Cornus officinalis fruits and their effects on diabetic nephropathyScienceDirect: S0378874114002414
  • Inhibition by Female Sex Hormones of Collagen Gel Contraction Mediated by Retinal Pigment Epithelial CellsPubmed: 24609629
  • Mesenchymal stromal cell proliferation, gene expression and protein production in human platelet-rich plasma-supplemented mediaPubmed:Pmc4130592
  • Metabolic and cytoprotective effects of in vivo peri-patellar hyaluronic acid injections in cultured tenocytesPubmed:25333747
  • Establishing principles of macromolecular crowding for in vitro fibrosis research of the vocal fold lamina propriaPubmed:25545625
  • In vivo bioengineered ovarian tumors based on collagen, matrigel, alginate and agarose hydrogels: a comparative studyPubmed:25634132
  • In vivoPubMed: 25634132
  • Impact of Nigella Sativa, Omega-3 Fatty Acids and Chromium Picolinate onNF-κB/leptin-insulin Axisin Obese Subjects with Non-alcoholic Fatty Liver DiseaseAjmbr: 3
  • Comparative proteomics of milk fat globule membrane in goat colostrum and mature milkPubmed:27173528
  • Maternal endothelial damage as a disorder shared by early preeclampsia, late preeclampsia and intrauterine growth restriction.pubmed:27865093
  • Effect of panax notoginseng saponins on efficacy and hemorrhagic transformation of rt-PA intravenous thrombolysis in patients with acute ischemic strokeArticle_en:CJFDTotal-XDJB201611014

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