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Multiplex Assay Kit for Amphiregulin (AREG) ,etc. by FLIA (Flow Luminescence Immunoassay)
AR; CRDGF; SDGF; Colorectum Cell-Derived Growth Factor; Schwannoma-Derived Growth Factor
(Note: Up to 8-plex in one testing reaction)
- Product No.LMA006Hu
- Organism SpeciesHomo sapiens (Human) Same name, Different species.
- Sample TypeSerum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
- Test MethodDouble-antibody Sandwich
- Assay Length4h
- Detection Range0.98-1000pg/mL
- SensitivityThe minimum detectable dose of this kit is typically less than 0.327 pg/mL.
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Result
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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.
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) | 83-101 | 90 |
| EDTA plasma(n=5) | 81-89 | 84 |
| heparin plasma(n=5) | 95-104 | 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) | 90-97% | 86-94% | 89-98% | 89-103% |
| EDTA plasma(n=5) | 88-96% | 80-95% | 89-97% | 78-93% |
| heparin plasma(n=5) | 89-97% | 94-105% | 95-104% | 86-101% |
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 |
| 96-well plate | 1 | Plate sealer for 96 wells | 4 |
| Pre-Mixed Standard | 2 | Standard Diluent | 1×20mL |
| Pre-Mixed Magnetic beads (22#:AREG) | 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. Add 200μL analysis buffer solution to each well of the plate for pre-wetting;
2. Preparation of standards, reagents and samples before the experiment;
3. Add 100μL standard or sample to each well, add 10μL magnetic beads, and incubate 120 minutes protect from light at 25°C on shaker;
4. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60 minutes protect from light at 25°C on shaker;
5. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent B, and incubate 30 minutes protect from light at 25°C on shaker;
6. Wash plate once on magnetic frame;
7. Add 100μL sheath fluid, swirl for 10 minutes, read on the machine.
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| Magazine | Citations |
| Onkologie | Amphiregulin and Epiregulin Expression in Colorectal Carcinoma and the Correlation with Clinicopathological Characteristics Karger: 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 Sci | Association between serum ligands and the skin toxicity of anti-epidermal growth factor receptor antibody in metastatic colorectal cancer PubMed: 25707609 |
| J Cancer Res Clin Oncol | Amphiregulin confers trastuzumab resistance via AKT and ERK activation in HER2-positive breast cancer PubMed: 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 cancer Pubmed:26716644 |
| Acta Pharmacologica Sinica | Anti-asthmatic activity of alkaloid compounds from Pericarpium Citri Reticulatae (Citrus reticulata 'Chachi') Pubmed: 19363516 |
| International Journal of Biological Sciences | CD9 regulates keratinocyte migration by negatively modulating the sheddase activity of ADAM17 |
| Neoplasia | Patient-derived xenograft (PDX) models of colorectal carcinoma (CRC) as a platform for chemosensitivity and biomarker analysis in personalized medicine Pubmed: 33212364 |
| JOURNAL OF CELLULAR AND MOLECULAR MEDICINE | Polarization of ADAM17‐driven EGFR signalling in electric field‐guided collective migration of epidermal sheets Pubmed: 33164313 |