Cell Proliferation, VEGF-Induced (Agonist & Antagonist), Panlabs
OVERVIEW
DOCUMENTS/REFERENCES
Biological Information
Background Information:
Vascular endothelial growth factor (VEGF), a homodimer of 34-42 kDa, has been detected in a wide range of normal and cancerous cells and upregulated by hypoxia, TGF-β1, phorbol ester and IL-6. The current nomenclature for the VEGF receptors lists three, entitled VEGFR-1 (Flt-1), VEGFR-2 (KDR/Flk-1) and VEGFR-3 (Flt-4). The receptors are transmembrane receptor tyrosine kinases and signal transduction through a phosphorylation cascade on initial ligand binding and autophosphorylation. VEGF function in vascular endothelial cells include induction of plasminogen activator and interstitial collagenase. Therefore, a specific VEGF stimulator or inhibitor may have potential clinical usage for different purposes.
No. of Assays on Panel:
2
Assays on Panel:
Family
Kinase
Name
Cell Proliferation, VEGF-Induced (Agonist), Panlabs
Item
307910-0
Protein
Organism
Human
Assay Sub Type
Cell Based
Family
Kinase
Name
Cell Proliferation, VEGF-Induced (Antagonist), Panlabs
Item
307910-1
Protein
Organism
Human
Assay Sub Type
Cell Based
Family:
Kinase
Sub Family:
RTK
Class:
Protein Tyrosine
Organism:
Human
Construct Details:
Endogenous
Cell Type:
HUVEC
Assay Information
Assay Type:
Functional
Assay Sub Type:
Cell Based
Functional Mode:
Agonist & Antagonist
Detection Method:
Luminescence
Measured Response:
Luminescence
Testing Information
Procedure Summary:
Human umbilical vein endothelial cells (HUVEC) are seeded in a 96-well plate and incubated at 37°C, 5% CO2 overnight. Test compound or vehicle is then incubated with cells at 37°C, 5% CO2 for 48 hours. After incubation, the reaction is evaluated by detecting ATP levels in living cells and the luminescence intensity is measured using luminescence reader. Test compound-induced stimulation of cell proliferation by 50 percent or more (≥50%), relative to 30 ng/ml VEGF165 control response, indicates possible agonist activity. Test compound-induced suppression of 2 ng/ml VEGF165-induced cell proliferation by 50 percent or more (≥50%) indicates possible antagonist activity.
Incubation:
2 d at 37°C
Control Inhibitor:
SU5416
Control Activator:
VEGF₁₆₅
Criteria For Significance:
≥ 50% Increase in proliferation relative to 30 ng/ml VEGF₁₆₅ response; ≥ 50% Inhibition of 2 ng/ml VEGF₁₆₅-induced proliferation
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
Additional Information
Brand:
Panlabs
Testing Location:
Taiwan - Taipei
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