Cultrex® In Vitro Angiogenesis Assay Tube Formation Kit

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	- Plates are coated with Reduced Growth Factor Basement Membrane Extract and observed by microscopy after 16h.

Protocol tips
- Plates are coated with Reduced Growth Factor Basement Membrane Extract and observed by microscopy after 16h.

Publication protocol

In vitro tube formation was assessed using The Cultrex In vitro Angiogenesis Assay Kit following manufacturer's protocol (Trevigen, Gaithersburg, MD). Cells were first counted, then PMVECs in starvation media containing either vehicle (H2O), BMP2 (10 ng/ml), or VEGF (50 ng/ml) were plated into 24-well plates coated with Reduced Growth Factor Basement Membrane Extract (Trevigen). For all experiments performed with WT and Tie2CrePPARγflox/flox mice, representative images of endothelial tube networks were obtained at 16 h under a fluorescence microscope, and the total tube area per field at 10× magnification was quantified in a blinded fashion using Bioquant Image Analysis software (R&M Biometrics, Nashville, TN). Calcein pre-treatment was used to fluorescently label all living murine PMVECs (Trevigen). In experiments performed with human PMVECs, cells were first fixed and stained following the protocol provided by the manufacturer (Trevigen) and the identification of endothelial tube formation was done under a light microscope (10× magnification) from three different fields. The total tube area per field at 10× magnification was quantified using Wimasis Image Analysis software (Wimasis GmbH, Munich, Germany).


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Manufacturer protocol

Download the product protocol from Bio-Techne for Cultrex® In Vitro Angiogenesis Assay Tube Formation Kit below.

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