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Cell adhesion assay

PE Pugazendhi Erusappan
JA Jahedul Alam
NL Ning Lu
CZ Cédric Zeltz
DG Donald Gullberg
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Forty-eight-well plates were coated with fibronectin (1 μg/cm2: Sigma-Aldrich) or collagen type I (5 μg/cm2: Bovine PureCol®, Advanced BioMatrix) and incubated for 2 hours at 37 °C. After washing with PBS, the plates were blocked with 2% BSA for 1 hour at 37 °C. Cells were washed three times with DMEM and 1 × 105 cells/well were cultured for 50 minutes at 37 °C. Unattached cells were removed carefully by washing three times with PBS containing Ca2+ and Mg2+. Cells were then fixed with 96% ethanol for 10 minutes at room temperature followed by staining with 0.1% crystal violet for 20 minutes at room temperature. Plates were washed three times with distilled water and the cells were lysed with 1% Triton X-100 for 5 minutes. The lysates were transferred to a 96-well plate and absorbance was read at 595 nm.

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