ELISA Kit for Amphiregulin (AREG) Mus musculus (Mouse) Sandwich ELISA

AR; CRDGF; SDGF; Colorectum Cell-Derived Growth Factor; Schwannoma-Derived Growth Factor

  • Product No.SEA006Mu
  • Organism SpeciesMus musculus (Mouse) Same name, Different species.
  • Sample Typeserum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
  • Test MethodDouble-antibody Sandwich
  • Assay Length3h
  • Detection Range15.6-1,000pg/mL
  • SensitivityThe minimum detectable dose of this kit is typically less than 5.8pg/mL.
  • Specificity This assay has high sensitivity and excellent specificity for detection of Amphiregulin (AREG).No significant cross-reactivity or interference between Amphiregulin (AREG) and analogues was observed.
  • ApplicationsEnzyme-linked immunosorbent assay for Antigen Detection.
  • DownloadInstruction Manual
  • UOM 48T96T 96T*5 96T*10 96T*100 Ensure in stock; if not, free redo!
  • FOB US$ 342.00 US$ 488.00 US$ 2,196.00 US$ 4,148.00 US$ 34,160.00
  • Quantity
  • Add to Cart Distributors
  • ELISA Kit for Amphiregulin (AREG) Packages (Simulation)
  • ELISA Kit for Amphiregulin (AREG) Packages (Simulation)
  • ELISA Kit for Amphiregulin (AREG) Results demonstration
  • SEA006Mu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 95-105 101
EDTA plasma(n=5) 79-103 81
heparin plasma(n=5) 93-104 99

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Amphiregulin (AREG) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Amphiregulin (AREG) 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 Amphiregulin (AREG) 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) 78-101% 84-97% 95-102% 95-105%
EDTA plasma(n=5) 88-99% 90-104% 83-101% 97-105%
heparin plasma(n=5) 88-97% 89-97% 79-101% 94-102%

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 Amphiregulin (AREG)

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

Reference

  • OnkologieAmphiregulin and Epiregulin Expression in Colorectal Carcinoma and the Correlation with Clinicopathological CharacteristicsKarger: 315380
  • Br J Cancer.Serum levels of hepatocyte growth factor and epiregulin are associated with the prognosis on anti-EGFR antibody treatment in KRAS wild-type metastatic colorectal cancer.Pubmed:24800946
  • Cancer SciAssociation between serum ligands and the skin toxicity of anti-epidermal growth factor receptor antibody in metastatic colorectal cancerPubMed: 25707609
  • J Cancer Res Clin OncolAmphiregulin confers trastuzumab resistance via AKT and ERK activation in HER2-positive breast cancerPubMed: 26195282
  • Oncotarget. Serum level of hepatocyte growth factor is a novel marker of predicting theoutcome and resistance to the treatment with trastuzumab in HER2-positive patients with metastatic gastric cancerPubmed:26716644

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