Chaim Sheba Medical Center View Institution's Website 5 articles published in JoVE Immunology and Infection T and B Cell Receptor Immune Repertoire Analysis using Next-generation Sequencing Lael Werner1,2, Chen Dor3,4, Naomi Salamon1, Meital Nagar3,4, Dror S. Shouval1,2 1Pediatric Gastroenterology Unit, Edmond and Lily Safra Children's Hospital, Sheba Medical Center, 2Sackler Faculty of Medicine, Tel Aviv University, 3Cancer Research Center, Sheba Medical Center, 4Molecular Hematology Laboratory, Sheba Medical Center The current protocol describes a method for DNA isolation from blood samples and intestinal biopsies, generation of TCRβ and IGH PCR libraries for next-generation sequencing, performance of a NGS run and basic data analysis. Immunology and Infection Co-immunoprecipitation Assay for Studying Functional Interactions Between Receptors and Enzymes Michael Peled1, Marianne Strazza2, Adam Mor2 1Pulmonary Department, The Chaim Sheba Medical Center, 2Columbia Center for Translational Immunology, Department of Medicine, Columbia University Medical Center Here, we present a protocol for co-immunoprecipitation and an on-bead enzymatic activity assay to simultaneously study the contribution of specific protein domains of plasma membrane receptors to both enzyme recruitment and enzyme activity. Biology Guided Protocol for Fecal Microbial Characterization by 16S rRNA-Amplicon Sequencing Ayelet Di Segni1, Tzipi Braun1, Marina BenShoshan1, Sarit Farage Barhom1, Efrat Glick Saar1, Karen Cesarkas1, James E. Squires2, Nathan Keller1,3, Yael Haberman1,4 1Sheba Medical Center, 2 This manuscript describes a detailed standardized protocol of high-throughput 16S rRNA-amplicon sequencing. The protocol introduces an integrated, uniformed, feasible, and inexpensive protocol starting from fecal sample collection through data analyses. This protocol enables analysis of large numbers of samples with rigorous standards and several controls. Medicine Cell Death Associated with Abnormal Mitosis Observed by Confocal Imaging in Live Cancer Cells Asher Castiel1, Leonid Visochek2,3, Leonid Mittelman4, Yael Zilberstein4, Francoise Dantzer5, Shai Izraeli1,6, Malka Cohen-Armon2,3 1Cancer Research Center, Sheba Medical Center, 2The Neufeld Cardiac Research Institute, Tel-Aviv University, 3Department of Physiology and Pharmacology, Tel-Aviv University, 4Imaging Unit, Sackler Faculty of Medicine, Tel-Aviv University, 5Biotechnology and Cell Signaling, Ecole Superieure de Biotechnologie Strasbourg, 6Department of Human Molecular Genetics and Biochemistry, Tel-Aviv University The cytotoxic activity of the phenanthridine PJ-34 in cancer cells undergoing mitosis was documented in real time by live confocal imaging. PJ-34 eradicated human breast cancer MDA-MB-231 cells harboring extra-centrosomes in mitosis. Unlike normal bi-focal mitosis, the extra-centrosomes were not clustered in the two spindle poles in the presence of PJ-34. Biology Ex Vivo Culture of Primary Human Fallopian Tube Epithelial Cells Susan Fotheringham1,2, Keren Levanon1,2,3, Ronny Drapkin1,2,4 1Department of Medical Oncology, Dana-Farber Cancer Institute, 2Harvard Medical School, Boston, MA, 3Sheba Cancer Research Center, Chaim Sheba Medical Center, 4Department of Pathology, Brigham and Women's Hospital The fallopian tube (FT) is emerging as an alternative site of origin for serous ovarian carcinoma (SOC). This protocol describes a novel method for the isolation and ex vivo culture of fallopian tube epithelial cells. This system recapitulates the in vivo epithelium and allows the study of SOC pathogenesis.