title: Immunocytochemical analysis of human pluripotent stem cells.
abstract: This chapter will describe the most common immunocytochemical method utilized in the stem cell field - using fluorescently tagged secondary antibodies to detect a primary antibody that is bound to an epitope on a molecule of interest. Secondary antibodies recognize the heavy chain of the primary antibody's isotype. Generally, these methods employ an incubation period of the sample with the primary antibody, a series of washes to remove unbound primary antibody, a secondary incubation period of the sample with the fluorescently conjugated secondary antibody, followed by washes and preparation for microscopy.
authors.affiliation: Neuroscience Laboratories, Centers for Neuroscience and Translational Research, CHOC Research Institute, Children's Hospital of Orange County Research Institute, Orange, CA, USA.
authors.forename: Hubert E
authors.initials: HE
authors.lastname: Nethercott
authors.forename: David J
authors.initials: DJ
authors.lastname: Brick
authors.forename: Philip H
authors.initials: PH
authors.lastname: Schwartz
mesh_terms: D001973:Bromodeoxyuridine / Q000378:metabolism; D049109:Cell Proliferation; D005456:Fluorescent Dyes / Q000378:metabolism; D005898:Glass; D006801:Humans; D021621:Imaging, Three-Dimensional; D007150:Immunohistochemistry / Q000379:methods*; D039904:Pluripotent Stem Cells / Q000737:chemistry / Q000166:cytology / Q000276:immunology / Q000378:metabolism*; D012984:Software; D013194:Staining and Labeling; D013499:Surface Properties; D016707:Tissue Fixation
chemical_list: D005456:Fluorescent Dyes; D001973:Bromodeoxyuridine