Cell Comb™ Scratch Assay

Protocol tips

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	- After washing cells they are incubated in 10 ml of FSM without or with inhibitors for 1-2 h.

Protocol tips
- After washing cells they are incubated in 10 ml of FSM without or with inhibitors for 1-2 h.

Publication protocol

"Cell scratches were carried out using Cell Comb Scratch Assay. 3T3-L1 preadipocytes plated in rectangular 1-well plates were grown in FSM to confluence. After removing the medium, plates were scratched in one direction with the Cell Comb. Cells were then washed once with DMEM to remove detached cells, and incubated in 10 ml of FSM without or with inhibitors for indicated period. Cells were also incubated for 1–2 h with inhibitors, where indicated, prior to scratching. Images were captured immediately after scratching and at indicated time after scratching using a light microscope equipped with camera (Canon, EOS 600D) and cell scratch areas were quantified using ImageJ software. Cell migration in this assay is presented as percentage closure of cell scratch area, calculated using the same equation that used for Oris cell
migration assay. Cells were then lysed as described in the ARF6-GTP pulldown assay and used to assess ARF6 activation and ERK1/2 phosphorylation"

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Manufacturer protocol

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