Does anyone have guidance on coating 96-well plates with collagen-I? Looking for a protocol. We are using Corning® Collagen I, Rat Tail, 100 mg.
Hello Jacob Leighty,
Use the following protocol.
1. Determine the volume of collagen-I solution needed.
2. Dilute collagen-I to 50ug/ml in 0.02M acetic acid to the needed volume.
3. Add diluted collagen at 5ug/cm^2 surface area. The area of a well for a 96- well plate is 0.33 cm^2. To coat 5ug/cm^2 of collagen-I, you will need 1.65ug of collagen-I per well in a 96 well plate i.e.,(5 x 0.33/1=1.65ug). You will use 50ul volume to cover a well of a 96 well plate.Therefore, you will need 1.65ug/50ul for each well.
4. Incubate for 1 hour at room temperature.
5. Carefully aspirate the remaining solution.
6. Rinse well 3 times to remove acid, using PBS or serum-free medium.
7. The plate may be used immediately or air dried and stored at 2-8° C for up to one week under sterile conditions.
Best.
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