Cell migration / Invasion cell type - Detroit 562

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 1 matching solution for this experiment

Protocol tips
Add 300 µL of prepared cell suspension from step 3 to each insert.

Add 500 µL of serum free media in the presence or absence of chemo-attractant (e.g. 10% fetal bovine serum) to the lower chamber.

Cover plate and incubate for 24 hours at 37°C in a CO2 incubator (4-6% CO2)
Downstream tips
Measure the Optical Density at 560 nm.
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