Detection of p24 in HIV-1 infected cells embedded in LR White and Lowicryl K4M.
In this study we present a postembedding on-grid immunogold labelling procedure for the ultrastructural localization of the HIV-1 core protein p24. HIV-1 infected cells were fixed in 0.1% glutaraldehyde, incompletely dehydrated and embedded in LR White or in Lowicryl K4M. Antigenic sites were detected by incubation of ultrathin sections with primary mouse monoclonal antibody anti-HIV-1 p24, followed by the secondary antibody goat anti-mouse IgG coupled to 10nm gold particles. Antigenicity of p24 was found to withstand the applied fixation and was shown to be preserved in LR White as well as in Lowicryl. The described procedure permits the uncomplicated and easy detection of p24 in HIV-1 infected cells and tissues.
Department of Medicine, University of Alabama, Birmingham 35294.
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D000180:Acrylic Resins; D000911:Antibodies, Monoclonal; D002460:Cell Line / Q000648:ultrastructure; D006046:Gold; D015488:HIV Antigens / Q000032:analysis*; D016655:HIV Core Protein p24 / Q000032:analysis*; D015658:HIV Infections / Q000378:metabolism / Q000382:microbiology*; D015497:HIV-1 / Q000737:chemistry / Q000302:isolation & purification*; D006801:Humans; D007150:Immunohistochemistry; D016253:Microscopy, Immunoelectron; D008867:Microtomy / Q000379:methods
D000180:Acrylic Resins; D000911:Antibodies, Monoclonal; D015488:HIV Antigens; D016655:HIV Core Protein p24; D006046:Gold; C037883:Lowicryl 4KM resin; C048707:LR white