Cd34 pe antibody

Cells derived from normal pancreas, chronic pancreatitis and pancreatic cancer tissues were analyzed on a flow cytometer (Beckman Coulter, Krefeld, Germany) with a software package (Coulter, Epics XL-MCL, System II). TLR7 antibody was purchased from Imgenex, TLR8 was provided by ProSci. CD34-PE antibody, FITC-conjugated anti-rabbit secondary antibody and isotype control antibodies were purchased by Beckman Coulter. For intracellular staining we used IntraPrep kit (Beckman Coulter).

NALM6^CBR-GFP or Ramos^CBR-GFP target cells were seeded at a density of 25,000 cells per well in a 96-well plate in RPMI complete media. UCART19 cells were purified from the spleens of NSG mice on day +28 by labeling with CD34-PE antibody (Beckman Coulter, IM1459U) followed by anti-PE microbeads (Miltenyi, 130-048-801) and selecting bead-bound cells on the AutoMACS. Cryopreserved UCART19 input cells from Day 0 were used as a control. UCART19 cells were co-cultured with target cells at E:T ratios ranging from 4:1 to 1:4 at 37 °C for 20 h. Absolute cell counts of viable target cells were quantified by flow cytometry using 7-aminoactinomycin D and GFP.