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Direct Staining of Intracellular Antigens by Flow Cytometry: the Paraformaldehyde/Saponin Method
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Alternative protocol for cell permeabilization required prior to intracellular staining, that does not require the use of Leucoperm
The detection of intracellular antigens requires a cell permeabilization step prior to staining. This method provides an alternative procedure for use when protocol FC7. “Direct Staining of Intracellular Antigens and Cytokines by Flow Cytometry using Leucoperm” does not provide the desired results. This method is particularly suitable for use with directly conjugated FITC monoclonal antibodies. In some cases specific recommendations are provided on product datasheets, and these methods should always be used in conjunction with the product and batch specific information provided with each vial. A certain level of technical skill and immunological knowledge is required for the successful design and implementation of these techniques; these are guidelines only and may need to be adjusted for particular applications.