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Direct Staining of Intracellular Antigens by Flow Cytometry: Methanol plus Leucoperm Method
Alternative protocol for cell permeabilization required prior to intracellular staining
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 by Flow Cytometry using Leucoperm does not provide the desired results. This method is particularly suitable for the detection of nuclear antigens, such as PCNA and Ki67. 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. Specific methodology for blood appears in [ ] brackets.