Transwell Invasion Assay procedure. Cells are seeded into the apical chamber of the transwell system FluoroBlok. The basal chamber is filled with cell culture media supplemented with fetal calf serum that serves as a chemoattractant. The membrane separating the chambers is coated with extracellular matrix proteins. Within 24-hours incubation in the presence or absence of test compounds, cells invade the basal chamber through the membrane and are quantified via Calcein staining.
Example of the Invasion Assay with MDA-MB-231 cells. 6.0x104 cells were added into each well of the tumor invasion system. Cells were treated with Gefitinib for 24-hours in the presence of a chemoattractant (20% FBS). The invaded cells were then stained with Calcein dye and fluorescence cells were read at wavelengths of 494/517 nm (Ex/Em). Data was acquired with Synergy Gen5 software and percent invasion was calculated using GraphPad Prism.
Setup: IC50 value determination with the BD Falcon HTS FluoroBlok 96-Multiwell insert system. Drug combination treatment is possible.
Controls: The DMSO only reactions are used as full activity control. Standard control compounds are tested in 8-dose IC50 format.
Assay development: Custom-tailored assays can be performed. This assay can be established with new cell lines according to the client’s research needs.
Report: Raw data, IC50 values, and curve fitting will be delivered in Excel format via email. Assay conditions, target, and substrate information are available upon request. Requirements for this information should be noted prior to study commencement.
Turnaround time: Results will be available within 3 weeks. Expedited scheduling and data delivery can be arranged prior to the commencement of the studies.
Screening facility: The assay is performed in Malvern, PA, USA.