OVERVIEW
DOCUMENTS/REFERENCES
Biological Information
Background Information:
IL-4 is the regulator of lymphocyte functions (Th2 differentiation and B-cell IgG1 and IgE class switch) to play an important role in the development of immune disorders, particularly allergies, and some autoimmune diseases. IL-4 signals through type I or type II IL-4 receptors. IL4 binds IL-4α, and then IL-4/IL-4Rα binds the IL-13Rα1 chain, generating the type II IL-4 receptor, mainly on non-hematopoietic cells. The formation of the type II IL-4 receptor complex induces JAKs activation, then STAT6 phosphorylation and translocation to the nucleus to bind specific DNA sequences. These are key players in the inflammatory response triggered either by an invading parasite or allergen. The development of monoclonal antibodies targeting IL-4Rα and IL-4 could be a clinical strategy for treating 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:
Agonist
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. Cells were incubated overnight and followed by test compound and/or vehicle treatment 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 increase of SEAP levels by 50 percent or more (≥50%) relative to the 10 ng/mL human IL-4 response indicates possible IL-4 receptor agonist activity. Test compound-induced inhibition of 0.2 ng/mL human IL-4 induced SEAP levels by 50 percent or more (≥50%) indicates IL-4 receptor antagonist activity
Incubation:
24 h at 37°C
Control Activator:
human IL-4
Criteria For Significance:
≥ 50% Increase in absorbance at 655 nm relative to human IL-4 response
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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