Epcam apc efluor780 clone g8.8

Tumors and 1 cm² of jejunum from Cre-negative, VA, and VAK mice were cut into small pieces using the McIlwain Tissue Chopper and digested on the gentleMACS Octo Dissociator with Heaters (program, 37C_m_TDK_1) using the mouse Tumor Dissociation Kit (Miltenyi Biotec 130–096–730) according to the manufacturer's instructions, and prepared cells were resuspended in 0.5% BSA in PBS. Cells were stained in the brilliant stain buffer (BD Biosciences 566349) containing antibodies for 30 minutes at 4°C in the dark. The following antibodies were used: CD19-APC-eFluor780 (clone 1D3; 1:400; eBioscience 47–0193–82), CD3ε-BV650 (clone 17A2; 1:100; BioLegend 100229), CD8α-BUV805 (clone 53–6.7; 1:50; BD Biosciences 564920), EpCAM-APC-eFluor780 (clone G8.8; 1:100; eBioscience 47–5791–82), and TCRδ-FITC (clone GL3; 1:200; eBioscience 11–5711–85). Dead cells were identified with Zombie NIR Fixable Viability dye (BioLegend 423106). Cells were acquired using a 5-laser BD LSRFortessa flow cytometer with DIVA software (BD Biosciences). Data were analyzed using FlowJo Software (RRID:SCR_008520) version 9.9.6.