I am trying to screen HEK293T cells by immunofluorescence and even though I coat the 96 well plates with poly-l-lysine (pll)after multipe washes (being very careful) there are few cells left. Has anyone fround anything better than pll?
Hello Angela Woods
When you culture HEK293T cells, you need to keep tight control over temperature. In case of HEK293T, cellular attachment is temperature dependent. It is imperative to use pre-warmed medium and reagents when working with these cells. If HEK293T cells are observed to detach from the substratum, this may not indicate a problem with viability, it may simply be the case that at some point the culture might have been at a temperature below 37°C for a short while.
If the cells become detached it is important to check cellular viability, and you need to wait for several days for the cells to re-attach. If these cells are required for cell based assay it is recommended that coatings such as Poly-D-Lysine (PDL) or collagen be used. You may also consider using CellBind (Corning product) to increase cellular attachment. Please refer to the link below for the product.
https://www.corning.com/in/en/products/life-sciences/products/surfaces/cell-bind.html
Good Luck!
Malcolm Nobre thanks for your reply. I have no problem with cell detachement when culturing cellsthe cells detach after fixation and many washes during immunofluorescent staining (even when really careful. I am going to try Cellbind plates and poly-D-lysine. thanks again
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