Eccles:Collagen Coating of Cell Culture Vessels: Difference between revisions
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| Line 11: | Line 11: | ||
!Flask/Well Surface Area (cm<sup>2</sup>) | !Flask/Well Surface Area (cm<sup>2</sup>) | ||
!Media Volume | !Media Volume | ||
!Volume Collagen Solution (0. | !Volume Collagen Solution (0.05ug/ul) to Achieve ~6ug/cm<sup>2</sup> | ||
|- | |- | ||
!96 well plate | !96 well plate | ||
| Line 28: | Line 28: | ||
|75||20ml||8.824ml | |75||20ml||8.824ml | ||
|} | |} | ||
====Collagen Solution (0.051ug/ul)==== | ====Collagen Solution (0.051ug/ul)==== | ||
Revision as of 16:10, 4 October 2007
Collagen Coating of Cell Culture Vessels
- Pipette collagen solution (0.051ug/ul collagen in 1x PBS) onto surface and swirl to disperse as you would with cells.
- Top up the volume with more 1x PBS if required (96, 6 wells and T75 flasks) so that the surface remains covered with liquid for the whole 2h.
- Leave collagen solution on plate surface for 2h (can be left in cell culture hood or incubator).
- Aspirate off liquid and rinse 2x in 1x PBS. Add media to keep collagen moist. Add cells as usual or vessel can be kept in cell culture incubator until ready to add cells.
| Vessel Size | Flask/Well Surface Area (cm2) | Media Volume | Volume Collagen Solution (0.05ug/ul) to Achieve ~6ug/cm2 |
|---|---|---|---|
| 96 well plate | 0.28 | 0.2ml | 33ul |
| 24 well plate | 2.0 | 0.5ml | 300ul |
| 6 well plate or 35x10mm round dishes | 9.6 | 2ml | 1.129ml |
| T25 flask | 25 | 5ml | 2.941ml |
| T75 flask | 75 | 20ml | 8.824ml |
Collagen Solution (0.051ug/ul)
- Stock collagen is at 1ug/ul
- Dilute collagen in 1x PBS to 0.051ug/ul ready for use (store at 4 degrees C)
| Collagen Stock (1ug/ul) | 1x PBS | |
|---|---|---|
| Volume (to make 50ml) | 2.55ml | 47.45ml |
