I used Poly-D-lysine for T cells and worked well in Seahorse mito stress. However, you need to normalized your experiment result by protein conc. of corresponding wells.

Cell-Tal works very well, but is more expensive than poly-D-lysine.

Thank you all.

Dear Gurcharan:

Usually for T and B cells, you do not even need cell-tak or poly-lys to do Seahorse. One way that works for me is just spinning them down in the Seahorse plate and then Mito-Stress. For a 1-2 h experiment will be just great.

Thanks Marcos.

But the seahorse protocol recommends you to plate the cells in 50ul of assay medium, spinning it and then adding more media after a few minutes.

If the cells are not attached and just centrifuged, wont the cells detach on addition of more media?

Yes.... but believe me. Cell tak is not need it. Cells will stick to the plate and form a monolayer. You just need to gently add media from the side of the wells. That is how we do it here for T & B cells. If you try and do not feel comfortable, I will recommend using Cell-Tak.

Hi Marcos

Thanks for the reply.

The problem I have been facing is that even after using cell tak, when I add more media to the plate (I do it gently from the side of the walls ) the cells still detach.

I am working with NK cells.

For lymphs we just centrifuge them and they'll stick to the plate. Have you tried to do that?

I use Poly-D-Lysine, it works fine and cheaper.