Mov18 zel antibody

The following mAbs were used for phenotypic analysis: APC-Cy7 Mouse Anti-Human CD3; FITC-anti-human CD4; APC-anti-human CD8; (BD Biosciences). Tumor cell surface expression of HER2 was detected by trastuzumab antibody (Herceptin, Genentech, San Francisco, CA), CD20 by biotinylated rituximab (Rituxan; Genentech, South San Francisco, CA, USA) followed by incubation with Strepavidin-APC. BsAb-IR expression was detected by Mov18/ZEL antibody (Enzo Life Sciences) followed by anti-mouse FITC or APC conjugated antibody (LifeBioscience). The isotype controls mIgG-APC-Cy7, mIgG-FIT, mIgG-APC, mIgG-PerCpCy5, mIgG-Pacific-Blue-A, and mIgG-PE-Cy7 were from eBiosciences and anti-mouse-AF488 from Invitrogen. Flow cytometric data were analyzed by FlowJo software. For intracellular staining, cells were permeablized using the FoxP3 staining buffer set (eBioscience), according to manufacturer’s instructions and labeled with: anti-human IFNg-PacificBlue, anti-human IFNg-PerCpCy5, anti-human TNFa-APC, anti-human IL2-PeCy7 (BD Biosciences).

The following mAbs were used for phenotypic analysis: i. biotin-SP-conjugated AffiniPure rabbit anti-human IgG (H+L) and ii. biotin-SP-conjugated AffiniPure rabbit anti-mouse IgG (H+L) (Jackson, West Grove, PA); iii. biotin-conjugated recombinant human FOLR1 (RD Systems, Minneapolis, MN / Thermo Scientific, Rockford, IL); i.v. streptavidin-APC (BD, San Jose, CA); v. BD ViaProbe (7-AAD); vii. anti-human CD3-FITC, anti-human CD4-FITC, anti-human CD8-FITC (eBioscience). In adoptive immunotherapy experiments, T cells from peripheral blood were obtained via retro-orbital bleeding and stained for the presence of human CD45, CD3 and CD8 using anti-human CD45-PE, anti-human CD3-PerCP/Cy5.5 and anti-human CD8-APC (Biolegend, San Diego, CA). After gating on the human CD45⁺ population, the CD3⁺ and CD8⁺ subsets were quantified using TruCount tubes (BD Biosciences) with known numbers of fluorescent beads as described in the manufacturer's instructions. Tumor cell surface expression of FR was detected by Mov18/ZEL antibody (Enzo Life Sciences). Flow samples were run using BD FACS Canto and cytometric data analyzed by FlowJo 7.6.5 software.