EXCELLYSE XPerm staining protocol

   
Procedure for intracellular staining with EXCELLYSE XPerm described bellow provides optimal results for most antigens analysed in clinical laboratory.
 
1. Pipette 100 µl of anticoagulated human blood into flow cytometry tube (12x 75 mm) containing appropriate antibody reagents (SURFACE MARKERS STAINING STEP)
 
2. Add 250 µl of XPerm Solution A. Mix and incubate for 10 minutes at room temperature (FIXATION STEP).
 
3. Add 3 ml of denionized water. Mix and incubate for 10 minutes at room temperature (RBC LYSIS STEP).
 
4. Centrifuge the cells at 300 x g for 5 minutes, decant the supernatant.
 
5. Add 250 µl of XPerm Solution B. Add appropriate intracellular antibody reagents. Mix and incubate for 15 - 30 minutes at room temperature in the dark (INTRACELLULAR STAINING STEP).
 
6. Wash by adding 2 ml of PBS, centrifuge the cells at 300 x g for 5 minutes, decant supernatant. Resuspend cells in 0.2 – 0.3 ml of PBS with 1% formaldehyde (WASHING AND FIXATION STEP). ALOT-Permeabilizace-Layout-1-1.jpg