ELISA Kit for Epiregulin (EREG)

EPR; ER; Proepiregulin

Specificity

This assay has high sensitivity and excellent specificity for detection of Epiregulin (EREG).
No significant cross-reactivity or interference between Epiregulin (EREG) and analogues was observed.

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 82-96 85
EDTA plasma(n=5) 99-105 102
heparin plasma(n=5) 83-96 92

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Epiregulin (EREG) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Epiregulin (EREG) 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 Epiregulin (EREG) 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) 80-98% 83-90% 93-102% 96-104%
EDTA plasma(n=5) 92-101% 81-89% 79-94% 97-104%
heparin plasma(n=5) 90-99% 96-103% 93-101% 90-104%

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.

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Magazine Citations
British Journal of Cancer Cross-suppression of EGFR ligands amphiregulin and epiregulin and de-repression of FGFR3 signalling contribute to cetuximab resistance in wild-type KRAS tumour cells Nature: 2012103
J Pharmacol Exp Ther. Aryl hydrocarbon receptor antagonism attenuates growth factor expression, proliferation, and migration in fibroblast-like synoviocytes from patients with rheumatoid arthritis. Pubmed: 24309559
Molecular Carcinogenesis The Ah receptor regulates growth factor expression in head and neck squamous cell carcinoma cell lines Pubmed:23625689
Am J Physiol Gastrointest Liver Physiol. Epiregulin promotes the emergence and proliferation of adult liver progenitor cells. Pubmed:24812054
European Review for Medical and Pharmacological Sciences Epiregulin-loaded PLGA nanoparticles increase human keratinocytes proliferation: preliminary data pubmed:27383296
Oncotarget Overexpression of a functional calcium-sensing receptor dramatically increases osteolytic potential of MDA-MB-231 cells in a mouse model of bone metastasis through epiregulin-mediated osteoprotegerin downregulation PMC5593575
American journal of respiratory cell and molecular biology Potential Involvement of the Epidermal Growth Factor Receptor Ligand Epiregulin and Matrix Metalloproteinase-1 in Pathogenesis of Chronic Rhinosinusitis pubmed:28398769
International Journal of Cancer Epigenetic regulation of Amphiregulin and Epiregulin in colorectal cancer Pubmed: 30252132
Epiregulin is released from intervertebral disks and induces spontaneous activity in pain pathways Doi: 10.1097/PR9.0000000000000718
Cell proliferation Epiregulin enhances odontoblastic differentiation of dental pulp stem cells via activating MAPK signalling pathway Pubmed: 31454111
International Journal of Molecular Sciences Promising Therapeutic Efficacy of GC1118, an Anti-EGFR Antibody, against KRAS Mutation-Driven Colorectal Cancer Patient-Derived Xenografts Pubmed: 31771279
Neoplasia Patient-derived xenograft (PDX) models of colorectal carcinoma (CRC) as a platform for chemosensitivity and biomarker analysis in personalized medicine Pubmed: 33212364
Reprod Sci Compromised Cumulus-Oocyte Complex Matrix Organization and Expansion in Women with PCOS 34748173
Tissue and Cell MiR-192-5p suppresses M1 macrophage polarization via epiregulin (EREG) downregulation in gouty arthritis 34715618
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