The amount of solution should cover the entire surface of the culture ware. In the case of 75cm^2 and 175cm^2, we recommend using a minimum of 5ml and 10ml respectively.

For a T75 flask, we recommend using 5ml - 7ml of our Applied Cell Extracellular Matrix (G422).

We suggest using 0.5 - 1ml to coat a T25 flask

You may use fibronectin coated flasks, however we cannot guarantee the cells will meet their maximum proliferation rate when doing so. The information specified for passage number and proliferation rate has been based on culturing these cells in ECM coated plates or flasks (Cat.No.G422)

We advise leaving the wells or dishes for ~1 hour to allow the coated surface to dry completely. Coated plasticware can then be used immediately without needing to wash with PBS. Alternatively, the coated flasks can be 4 - 8ºC for up to 2 weeks.

It can be used on both.

We recommend using 20mL of G422 to coat a T175 flask. You may top up the volume using 1X sterile PBS to keep collagen moist while coating. This can be done by diluting G422 (1:1) with PBS and follow the rest of the protocol as per our website.

This can occur if the ECM is used in excess or if it is not coated evenly. We suggest diluting the ECM with media (1:1) and leaving it for a longer time to produce a more homogeneous layer.

We have had success in creating a thin coating by diluting the G422 with PBS before coating. Additionally, it has been found that shorter incubation time and dilution with media has also helped.

For a dish that is 100mm x 20mm (i.e. with a surface area that is roughly ~55 cm^2), we would recommend ~1.5-2 mL of matrix to cover the surface. However, your are certainly able to add a bit more of the matrix if you are looking for a thick coating for your application.

G422 may have some lipid traces or related materials since it is complex heterogeneous mixture of extracellular matrix.