What positive control for intracellular staining? - (Nov/28/2011 )
I need to learn well technique of intracellular staining of PBMC on flow cytometry in order to make sure that later, when I do not see protein of my interest, the reason is that this protein is not expressed by the cell not because I am not able to make the procedure the correct way. But, I do not know what primary antibody, against what protein to buy and use. What intracellular protein I shall use to train detection by FCM to be 100% positive it is always expressed in PBMC...?
When people need to ensure their cells are properly permeabilized and in good condition, they'll sometimes use an antibody to F-Actin, which is present in all cells. If the F-Actin staining is always positive then at least you know that the cells are properly permeabilized and that your buffers are working properly. This could serve as a positive control.