ELISA Kit for Tenascin C (TNC) Rattus norvegicus (Rat) Sandwich ELISA

TN-C; HXB; TN; GMEM; JI; Myotendinous antigen; Neuronectin; GP 150-225; Cytotactin; Hexabrachion; Glioma-associated-extracellular matrix antigen

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  • ELISA Kit for Tenascin C (TNC) Packages (Simulation)
  • ELISA Kit for Tenascin C (TNC) Packages (Simulation)
  • ELISA Kit for Tenascin C (TNC) Results demonstration
  • SEB975Ra.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 85-103 90
EDTA plasma(n=5) 90-97 94
heparin plasma(n=5) 84-101 93

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Tenascin C (TNC) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Tenascin C (TNC) 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 Tenascin C (TNC) 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) 78-103% 86-95% 78-99% 91-101%
EDTA plasma(n=5) 95-103% 95-102% 98-105% 96-104%
heparin plasma(n=5) 98-105% 91-105% 83-104% 95-102%

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 Tenascin C (TNC)

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

Citations

  • An investigation of tenascin-C levels in rheumatic mitral stenosis and their response to percutaneous mitral balloon valvuloplastyPubmed: 22889719
  • Clinical significance of serum tenascin-c levels in epithelial ovarian cancerSpringer: Source
  • Clinical significance of serum tenascin-C levels in breast cancerSpringer: Source
  • Tenascin-C is increased in atherothrombotic stroke patients and has an anti-inflammatory effect in the human carotid artery.Pubmed:24823872
  • Metabolic and cytoprotective effects of in vivo peri-patellar hyaluronic acid injections in cultured tenocytesPubmed:25333747
  • Role of Tenascin-C in the differential diagnosis of small round blue cell tumorsIndex.Php: Sdutfd
  • A new predictor of mortality in hemodialysis patients; Tenascin-CPubMed: 26390818
  • Biomarkers Score for Patients with Mitral Stenosis: A useful conjunction with Wilkins’s Score for Early InterventionHeartresearchopenjournal: Hroj5
  • Pankreas Adenokarsinomunda Serum Tenascin-C Düzeyi Potansiyel Bir Biyobelirteç Midir?Cpdf:5C21620161255
  • Increased tenascin C and DKK1 in vitiligo: possible role of fibroblasts in acral and non-acral diseasePubmed:29605863
  • How to detect and purify tenascin-CPubmed:29310788
  • Altered transcriptional regulatory proteins in glioblastoma and YBX1 as a potential regulator of tumor invasionPubmed: 31358880

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