scratch wound assay - (Jan/23/2008 )
Does anyone have a detailed and easy protocol for scratch wound assay?
1. Grow cells to monolayer
2. Take a yellow (p200) tip or smaller if you need
3. with even pressure scrape across surface in a straight line(be careful if your cells are on coverslips- if you push too hard you crack the glass)
4. If you want you can do multiple lines- say for protein extracts
5. Harvest or fix and stain cells for what ever analysis you want to do at what ever time points you want.
Thats the easiest protocol
cool! thats gr8! but do you do the scratch when the cells are still in the media. or you remove the media, wash with pbs and then perform it.
it is not easy to scratch with a tip; especially, if you have strongly coherent cells, the scratch may be (too) scruffy; you have to find your way to do the scratch...
Yep, Like The Bearer says it will depend on your cells- you could try both ways and see what works best for giving the most consistent results and straightest lines. When you wash the cells with PBS, I would do it gently though and try not to overgrow your cells or you might detach the whole sheet. Do a couple of practice ones to make sure your happy with your techniques and then off you go.