Cd5 phycoerythrincyanine 5.5 pc5.5

Intracellular staining of Ki-67 was performed using a fluorescein isothiocyanate (FITC)-labeled antibody against Ki-67 (Becton Dickinson, Franklin Lakes, NJ, USA) after fixation and permeabilization using the BD Intrasure kit (Becton Dickinson) following the manufacturer's instructions. Surface staining of cells was performed using the following monoclonal antibodies conjugated with the indicated fluorochrome: CD19-phycoerythrin (PE) and CD5-allophycocyanine (APC) (Becton Dickinson).
Expression of CD69, CD38 and CD86 in CD19⁺/CD5⁺ and CD3⁺ cells was assessed using the following antibodies: CD19-energy coupled dye (ECD), CD5-phycoerythrincyanine 5.5 (PC5.5) (Beckman Coulter, Brea, CA, USA), CD38-PE (EBioscience, San Diego, CA, USA), CD3-PE-cyanine 7 (Cy7), CD69-APC and CD86-APC (Becton Dickinson). Cells were acquired in a Navios^TM cytometer (Beckman Coulter, Brea, CA, USA) and the results were analyzed using the FCS Express 4 software (De Novo Software, Los Angeles, CA, USA).

Intracellular staining of Ki-67 was performed using a fluorescein isothiocyanate (FITC)-labeled antibody against Ki-67 (Becton Dickinson, Franklin Lakes, NJ, USA) after fixation and permeabilization using the BD Intrasure kit (Becton Dickinson) following the manufacturer's instructions. Surface staining of cells was performed using the following monoclonal antibodies conjugated with the indicated fluorochrome: CD19-phycoerythrin (PE) and CD5-allophycocyanine (APC) (Becton Dickinson). To characterize the phenotype of proliferative and resting compartments of CLL cells,we used the following antibodies: CD19-energy coupled dye (ECD), CD5-phycoerythrincyanine 5.5 (PC5.5) (Beckman Coulter, Brea, CA, USA), CD3-PE-cyanine 7 (Cy7), CXCR4-APC, CXCR5-APC, CCR7-APC, CD49d-APC, CD62L-APC, Ki-67-FITC (Becton Dickinson), and CD38-PE (EBioscience, San Diego, CA, USA). The rates of T cell activation and proliferation were analyzed by determining the expression of Ki-67, CD69 and CD38 in CD3+ cellsusing the following antibodies: Ki-67-FITC, CD38-PE (EBioscience), CD5-PC5.5 (Beckman Coulter), CD3-PE-Cy7 and CD69-APC (Becton Dickinson). Cells were acquired in a Navios^TM cytometer (Beckman Coulter) and the results were analyzed using the FCS Express 4 software (De Novo Software, Los Angeles, CA, USA).