Pe labeled goat anti mouse igg

Isolated T cells were stained with fluorochrome-labeled mabs directed against human CD4/VioBlue (Miltenyi Biotec, clone VIT4), CD3/PE-Cy7 (Biolegend, San Diego, USA, clone UCHT1), CD8/APC (BD Bioscience, clone RPA-T8), and CD25/PE (MiltenyiBiotec, clone 4E3). For detection of CAR surface expression T cells were incubated with mab 7B6 and subsequently stained with PE-labeled goat anti-mouse IgG (Beckmann Coulter, Krefeld, Germany). Samples were analyzed using a MACSQuant® Analyzer and MACSQuantify® software (Miltenyi Biotec) [47 ].

Isolated T cells were stained with fluorochrome-labeled mabs directed against human CD4/VioBlue (Miltenyi Biotec, clone VIT4), CD3/PE-Cy7 (Biolegend, San Diego, USA, clone UCHT1), CD8/APC (BD Bioscience, clone RPA-T8), CD27/PE (BD Bioscience, clone M-T271) and CD62L/PacificBlue (Biolegend, clone DREG-56). For detection of ECAR surface expression, T cells were incubated with anti-La mab 5B9 [16] and subsequently stained with PE-labeled goat anti-mouse IgG (Beckmann Coulter, Krefeld, Germany). Samples were analyzed using the MACSQuant Analyzer and the MACSQuantify software (Miltenyi Biotec). In order to assess expansion rates of ECAR armed T cells, absolute T cell numbers were quantified using a MACSQuant Analyzer and MACSQuantify software (Miltenyi Biotec) as described elsewhere [25] (link).

Isolated T cells were stained with fluorochrome-labeled mAbs directed against human CD4/FITC (clone VIT4, MiltenyiBiotec), CD3/Vioblue (MiltenyiBiotec) and CD8/APC (clone BW 135/80, MiltenyiBiotec). For analysis of expression of GD2 on JF cells, the cells were stained using the commercial anti-GD2 mAb (Biolegend, San Diego, USA; Clone 14G2a), detected with Alexa Flour 647-labeled goat anti-mouse IgG (Life technologies, Thermo Fisher Scientific). For detection of CAR surface expression T cells were incubated with the mAb 7B6 and subsequently stained with PE-labeled goat anti-mouse IgG (Beckmann Coulter, Krefeld, Germany). Samples were analyzed using a MACSQuant^® Analyzer and MACSQuantify^® software (Miltenyi Biotec) [56 (link)].