Excellent Permeabilization Buffer from eBioscience

For staining the intracellular antigens, cells need to be permeabilize before incubation with the antibody, so that antibody can easily penetrates into the cells and specifically binds to the target protein. We have used this permeabilization buffer for staining intracellular cytokines. Briefly splenocytes single cell suspension was prepared, RBCs were lysed using RBCs lysis buffer, cells were washed twice with facs buffer (1 X PBS + 2 % FBS). Fixed with fixation buffer for 30 minutes and washed twice with 1X permeabilization buffer. After that splenocytes were incubated with targeted cytokine specific antibody in permeabilization buffer for 1 h, washed twice with 1X permeabilization buffer and cell pellet was re-suspended in facs buffer for acquiring and analysis by flow cytometry.