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)

Specificity

This assay has high sensitivity and excellent specificity for detection of Amphiregulin (AREG) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Amphiregulin (AREG) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 78-101 90
EDTA plasma(n=5) 96-103 101
heparin plasma(n=5) 96-105 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Amphiregulin (AREG) ,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 Amphiregulin (AREG) ,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 Amphiregulin (AREG) ,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) 78-94% 79-94% 95-102% 89-98%
EDTA plasma(n=5) 98-105% 80-91% 82-98% 85-96%
heparin plasma(n=5) 93-101% 98-105% 93-101% 83-99%

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. 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.

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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
Catalog No. Related products for research use of Mus musculus (Mouse) Organism species Applications (RESEARCH USE ONLY!)
RPA006Mu02 Recombinant Amphiregulin (AREG) Positive Control; Immunogen; SDS-PAGE; WB.
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PAA006Mu01 Polyclonal Antibody to Amphiregulin (AREG) WB; IHC; ICC; IP.
PAA006Mu02 Polyclonal Antibody to Amphiregulin (AREG) WB; IHC; ICC; IP.
SEA006Mu ELISA Kit for Amphiregulin (AREG) Enzyme-linked immunosorbent assay for Antigen Detection.
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