Hi all,

I carried out a flow cytometry in adipose tissue with a new permeabilization and fixation method in order to improve the intracellular staining for TFs... However, comparing with the previous experiments (where I used a fixation and permeabilization method for membrane not nuclear) I realized that the number of cells is much lower... Therefore, I was wondering if this could be a technical issue related to the new kit. I used these reagents 00-5523-00 from Thermo Fischer .

I fixed the cells with the diluted fixation buffer for 45 min on ice.

Then I incubated with intracellular abs diluted in the permeabilization buffer (that was diluted 1:10 in water) for 1 hour on ice and then washed with the diluted permeabilization buffer.

Thanks!!

Rosa