IL4RA/IL13RA1 Human Interleukin Cell Based Antagonist IL-13-induced STAT6 Reporter LeadHunter Assay - TW
OVERVIEW
DOCUMENTS/REFERENCES
Biological Information
Background Information:
IL-13 is a cytokine that specifically regulates IgE synthesis, mucus secretion, and eosinophil functions during asthma and parasitic infections. IL-13 bridges allergic inflammatory cells to the non-immune cells in contact with them, thereby altering physiological function. The overexpression of IL-13 induces airway hyperresponsiveness, goblet cell metaplasia, mucus hypersecretion, subepithelial fibrosis, and airway remodeling, ultimately leading to airway obstruction. IL-13 directly binds IL-13Rα1, and the IL-13/IL-13Rα1 complex then recruits IL-4Rα into the functional receptor complex, type II IL-4 receptor, to activate the STAT6 transcription factor in non-hematopoietic cells, such as epithelial cells. Monoclonal antibody targeting interleukin-13 is the treatment for ulcerative colitis and atopic dermatitis. Monoclonal antibodies targeting IL-4Rα may be a treatment for asthmatic patients.
Family:
Immune System
Class:
IL-4
Organism:
Human
Construct Details:
Endogenous
Cell Type:
HEK293
Assay Information
Assay Type:
Functional
Assay Sub Type:
Cell Based
Functional Mode:
Antagonist
Detection Method:
Spectrophotometry
Platform:
Plate reader
Measured Response:
Absorbance
Testing Information
Procedure Summary:
HEK-Blue IL-4/IL-13 cells (2.5 x 10^5 /mL) were seeded in a 96-well plate. After incubation overnight, cells were treated with test compound and/or vehicle for 30 minutes and then stimulated by recombinant human IL-13 (2 ng/mL) at 37℃ with 5% CO2. After 24 h incubation, supernatants were harvested and mixed with reconstituted QUANTI-Blue substrate in a 96-well plate. The reaction is incubated at 37℃ for 1.5 h and the absorbance is read at 655 nm. Test compound-induced inhibition of 2 ng/mL human IL-13 induced SEAP levels by 50 percent or more (≥50%) indicates IL-13 receptor antagonist activity.
Incubation:
24 h at 37°C
Control Inhibitor:
Dupilumab
Control Activator:
human IL-13
Criteria For Significance:
≥ 50% inhibition relative to human IL-13-induced SEAP production
Test Concentration / Dose:
Compounds are screened at 10, 3, 1, 0.3, 0.1, 0.03, 0.01 and 0.003 μM
Test Sample Requirements:
75 μl of 10 mM stock or 1 mg (pre-weighed) powder (assuming FW ≦500 and purity 100%)
Minimum Order Quantity:
16
Turnaround Time
Standard:
20 Business Days
Reference Compound Data
Clinical Relevance
Therapeutic Area:
Inflammation/Allergy
Additional Information
Brand:
LeadHunter
Testing Location:
Taiwan - Taipei
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