Coating 24 well plates - (Nov/22/2006 )
Does anyone have a good protocol for coating 24 or 96 well plates with collagen or poly-L-lysine?
thanks a lot
I cover the well surface with some few microliters of Col-IV (25ug/ml) or p-L-Lys (stock) or Gelatin (0.1%). Incubate at RT on culture hood around 3-5 min. Recover the excess of liquid (I re-use it once, some people don't) and let the plate uncover dry completely. Then the plates are ready to use.
I cover surface of 12 well plate with 200 microliters of p-L-Lys (stock) . incubate for 2'h at co2 incubator . then i Recover the excess of liquid and dry completely. Then i seed my cell (PC12) for 24'h but i never got any cell attachement , why
we coat using poly-D lysine.
Make solution 100ug/ml and use it asd 10x and coat plates O/N at RT. next day dry it out and plate cells.
for 12 well plate i cover with collagen type I diluted with hydrochloric acid pH 3 10 folds , use 500 micron per well and leave it for one hour at RT then remove it by aspiration and wash the wells twice with media.