Cell migration / Invasion cell type - T47D

Cell Invasion or Cell Migration assays are technically challenging to set up as the gradient between the two compartments equilibrates in time during the assay. It is also problematic to view cells and for cells to migrate through a non-physiologic polycarbonate or polypropylene filter. Care must be taken while loading the well with cells to form a single cell suspension. Precaution must be taken while trypsinization (under-trypsinization can lead to cell clumping while over-trypsinization could strip off adhesion molecules necessary for migration). This leads to difficulty in getting significant results, when only small numbers of cells cross the filter or when the distribution and/or staining of the cells is uneven.

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Found 3 matching solutions for this experiment

Upstream tips
- Seed 5 × 10^5 cells/transwell.
Protocol tips
- Reconstitute Matrigel basement membrane with 5 μg/cm^2 .

- Supplement with 1% FBS medium on both sides of the wells and stiluli on the lower side.
Upstream tips
Seed 50,000 cells
Protocol tips
Incubate at 37oC in CO2 incubator for 24 hours
Downstream tips
Read assay chamber solutions (bottom) at 485 nm excitation, 520 nm emission
Upstream tips
Seed 3 × 10^5 cells
Protocol tips
Incubate for 24hours at 37ºC in 5% CO2 atmosphere.
Downstream tips
Measure the OD 560nm in a
plate reader
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