Angiogenesis, Tube Formation Assay in HUVEC Human Cells, Panlabs

Procedure Summary:

HUVECs (human umbilical venous endothelial cells, ATCC CRL-1730) are seeded in a 96-well plate pre-coated with basement membrane matrix. Test substance or vehicle is then incubated with the cells in incubation buffer under an atmosphere of 5% CO2 at 37°C. After an 18-hour incubation period, morphology of the endothelial cell tubes, which resemble a capillary-like network, are evaluated by ImageXpress Micro Confocal High-Content imaging system. Disruption (anti-angiogenesis) of total tube length is measured from each image and determined relative to the vehicle control group. IC50 value will be determined if a dose-response is available and the % inhibition includes values both above and below 50%.

Incubation:

1080 min at 37°C

Control Inhibitor:

Suramin

Criteria For Significance:

≥ 50% inhibition in total tube length relative to the vehicle control

Test Concentration / Dose:

Compounds are screened at 30, 3, 0.3, 0.03 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:

20