IntraPrepIntraPrep Permeabilization Reagent is a set of two ready-to-use reagents designed for the immunological detection of intracellular leucocyte antigens by flow cytometry. It uses a formaldehyde / saponin-based permeabilization procedure. IntraPrep Permeabilization Reagent can be used for detection of most cytoplasmic and nuclear antigens in whole blood samples. It creates apertures in the membrane without affecting the gross morphology of the cell. The resolution of each cell type on a dual light scatter histogram is well preserved during processing with this reagent system. The standard procedure consists of first, fixing cells with Reagent 1, and second, permeabilizing the white cells and lysing red cells using Reagent 2. The reagent formulation is optimized to minimize non-specific intracellular staining. The IntraPrep Permeabilization Reagent can be used for simultaneous detection of cell surface and cytoplasmic antigens by flow cytometry.
The PerFix-nc Kit (no centrifuge assay Kit) has been developed to enhance the signal-to-noise ratio of intracellular staining and simplify the workload necessary for the sample preparation. Accurate detection of both intracellular and extracellular epitopes are obtained, while:
- There are no washing steps through the whole STRAIGHT procedure (only a final wash step is described as optional).
- Several surface markers can be added together with the intracellular markers and incubated simultaneously during the permeabilization step.
- Total duration of the procedure and total workload are similar to current procedures for surface staining (e.g. VersaLyse + Fix No Wash), and much shorter than common permeabilization procedures.
- Potential automation of the PerFix-nc STRAIGHT procedure is rendered possible thanks to the removal of the washing steps.