Detection of p24 in HIV-1 infected cells embedded in LR White and Lowicryl K4M |
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| Authors: | Gregor Stransky Robert F Garry and Steffen Gay |
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| Institution: | (1) Division of Clinical Immunology and Rheumatology, Department of Medicine, The University of Alabama at Birmingham, 35294 Birmingham, Alabama, USA;(2) Department of Microbiology and Immunology, Tulane University School of Medicine, 70112 New Orleans, Louisians, USA |
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| Abstract: | Summary 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. |
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