FIX&PERM®

For each sample to be analyzed use 50 µl of whole blood, bone marrow or mononuclear cell suspension in a 5 ml tube

1) Add 100 µl of Reagent A (Fixation Medium, stored and used at room temperature).

2) Incubate for 15 minutes at room temperature. 

3) Add 5 ml phosphate buffered saline and centrifuge cells for 5 minutes at 300 g.

4) Remove supernatant and add to cell pellet 100 µl Reagent B (Permeabilization Medium) and 20 µl of the appropriate monoclonal antibody conjugate and/or the corresponding isotype controls.

5) Vortex at low speed for 1-2 seconds.

6) Incubate for 15 minutes at room temperature.

7) Wash cells with phosphate buffered saline as described above.

8) Remove supernatant and resuspend cells in sheath fluid for immediate analysis or resuspend cells in 0.5 ml 1.0 % formaldehyde and store at 2-8°C in the dark.

9) Analyze fixed cells within 24 hours.

 

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