how to coat 6-well plate with matrigel or laminin? - (May/17/2007 )

hi all
some cells(such as embryonic stem cell) grow well when seeded onto 6-well plate coated with laminin. but I don't know how to coat laminin.
for example, the protocol tells me to coat plate with 20ug/ml laminin, and now i have a laminin solution with a concentration of 20ug/ml, what should I next do?
just add 1ml(or other volume?) laminin solution in a well of a 6-well plate, and wait for all of the liquid evaporate to dryness? or I can aspirate the liquid after 1 hour or more?

Any suggestions are welcome.
thank you.

Our protocol goes something like this: apply a sufficient volume of the laminin at the desired concentration to each well ( we aim for 1-2 ug/cm2) and incubate at R.T. or 37 deg Celsius for 2 hours. Then aspirate and discard the remaining laminin. Allow the wells to air dry air dry at R.T. under a hood. If contamination is a possibility then air dry with the UV light on. Then rinse the wells several time with water or PBS and plate your cells or air dry and store at 4 deg Celsius for up to one month.

hope this helps and let me know if you have any other questions.

With matrigel, dilute it 1:40 in DMEM.

Use 150ul for 24 well plate (each well) or 500ul for a well in 6 well plate.

Add matrigel and make sure it covers the entire surface. Then immediately remove the matrigel. and let it dry for 30-40 min.


For Laminin,

coat plate with laminin, I use 250ul for 24 well plates. Let it sit for 1 -2 hrs and then wash it with media and seed the cells.

thank you very much
you help me so much