The Permeabilization Buffer (10X), in conjunction with the Intracellular (IC) Fixation Buffer (cat. 00-8222) or Foxp3 Transcription Factor Fixation/Permeabilization Concentrate and Diluent (cat. 00-5521), can be used to fix and permeabilize cells prior to performing intracellular staining of cytokines and other cytoplasmic and/or nuclear antigens. These solutions have been specially formulated for reducing non-specific staining of fluorochrome-labeled antibodies and increasing fluorescence signal to noise ratios.
In intracellular staining protocols, fixation is performed first, followed by permeabilization. Subsequent washing steps, antibody dilutions, and incubations after permeabilization should use the Permeabilization Buffer. The final washing step and cell resuspension prior to running the sample is done using Flow Cytometry Staining Buffer.
Intracellular Staining Followed by Flow Cytometric Analysis