Immuno-gold replica technique for ultrastructural localization of antigens | Immuno-gold replica technique for ultrastructural localization of antigens
1994
Hernández, Francisco | Rivera, Patricia | Hosaka, Yasuhiro
English. We developed a simple method for immunolabelling viral glycoproteins on surface of infected ceil monolayers. Cells 9 hours post-infected with Sendai virus were fixed and treated with polyclonal antibody anti HN glycoprotein (Rabbit IgG), labelled with goat immunoglobulin anti rabbit IgG conjugated with colloidal gold particles. Then, the cells were shadowed with platinum-carbon, covered with nitrocellulose glue, and digested with domestic bleach, washed, and replica fragments were caught with uncovered grid.
Show more [+] Less [-]Spanish; Castilian. Se describe un método simple para inmunomarcación de glicoproteínas virales en la superficie de células infectadas. Monocapas celulares 9 h post infección con virus Sendai fueron fijadas y tratadas con un anticuerpo policlonal (Conejo Ig6) anti la glicoproteína HN, marcado con inmunoglobulina de cabra anti IgG de conejo, conjugada con oro coloidal. Luego, las células fueron sombreadas con platino-carbón, cubiertas con nitrocelulosa y digeridas con blanqueador doméstico, lavadas y los fragmentos de réplicas fueron recuperados sobre rejillas sin membrana soporte.
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