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Direct staining of intracellular antigens and cytokines by Flow Cytometry: Leucoperm method
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Method for cell permeabilization required prior to intracellular staining using Leucoperm Accessory Reagent.
The detection of intracellular antigens requires a cell permeabilization step prior to staining. For the detection of cell cycle antigens such as PCNA and BrdU, methanol modification is recommended – see protocol FC8 (Direct staining of intracellular antigens by Flow Cytometry: Leucoperm plus methanol method).
The method described below produces excellent results in our hands; however, other permeabilization techniques have been published, and may also be used successfully for this application. 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. Note: Specific methodology for blood appears in [ ] brackets.