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URL: https://dx.doi.org/10.17504/protocols.io.bhbij2ke

Authors: Benjamin Izar, Parin Shah, Mei-Ju Su, Isaac Wakiro, Sara Napolitano, Jingyi Wu, Sébastien Vigneau, Asaf Rotem, Orit Rozenblatt-Rosen, Bruce Johnson, Aviv Regev

Group: NCIHTAN

Summary: Many ovarian cancer ascites samples have a high proportion of CD45-expressing cells (>90%) (CD45+) and a low proportion (For the Human Tumor Atlas Pilot Project (HTAPP) this protocol was used to deplete CD45+ immune cells from single-cell suspensions obtained from ovarian cancer ascites, and is included as a step in the "HTAPP_Processing human ovarian cancer ascites to a single-cell suspension for single-cell RNA-seq" protocol, also available in protocols.io. It is adapted from the CD45 MicroBeads MACS Separation protocol from Miltenyi Biotec (Cat No 130-045-801). Description of this protocol and guidance for testing and selecting methods for processing different tumor and sample types can be found in Slyper et al.

Affiliations: Dana-Farber Cancer Institute;Human Tumor Atlas Pilot Project, Dana-Farber Cancer Institute;Human Tumor Atlas Pilot Project, Dana-Farber Cancer Institute;Human Tumor Atlas Pilot Project, Dana-Farber Cancer Institute, University of California, Santa Barbara, Dana-Farber Cancer Institute;Human Tumor Atlas Pilot Project, Dana-Farber Cancer Institute;Human Tumor Atlas Pilot Project, Dana-Farber Cancer Institute;Human Tumor Atlas Pilot Project, Broad Institute;Human Tumor Atlas Pilot Project, Dana-Farber Cancer Institute;Human Tumor Atlas Pilot Project, Broad Institute;Human Tumor Atlas Project;Massachusetts Institute of Technology;Howard Hughes Medical Institute

Version: 2

Publication Date: 2020

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