Figure 2

Titration of Protein L concentration required for optimal FACS analysis. Activated PBLs were transduced with retroviral vector expressing anti-PSCA-CAR or anti-EGFRvIII-CAR and analyzed for CAR expression on day 8. One million cells were stained with 0.01, 0.05, 0.1, 1.0, 1.5, 2.0 or 3.0 μg/sample of biotinylated Protein L. The cells were then washed and stained with phycoerythrin (PE)-conjugated streptavidin (SA). Cells were analyzed using a FACScan flow cytometer and the data analyzed with FlowJo software. Result of a representative experiment from four independent experiments is presented. No-Prot-L, samples stained with SA-PE alone.