Immunohistochemical Detection of ZEBOV

A partial necropsy was performed on all subjects. Tissue samples of major organs were collected for histopathologic and immunohistochemical examination, immersion-fixed in 10% neutral buffered formalin, and processed for histopathology as previously described^{27 (link),28 (link)}. For immunohistochemistry, specific anti-ZEBOV immunoreactivity was detected using an anti-ZEBOV VP40 primary antibody (IBT) at a 1:4000 dilution for 60 min. The tissues were processed for immunohistochemistry using the Thermo Autostainer 360 (ThermoFisher, Kalamazoo, MI). Secondary used was biotinylated goat anti-rabbit IgG (Vector Labs, Burlingame, CA #BA-1000) at 1:200 for 30 min followed by Vector Horseradish Peroxidase Streptavidin, R.T.U (Vector) for 30 min. Slides were developed with Dako DAB chromagen (Dako, Carpenteria, CA #K3468) for 5 min and counterstained with Harris Hematoxylin for 30 s.

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Cross R.W., Bornholdt Z.A., Prasad A.N., Geisbert J.B., Borisevich V., Agans K.N., Deer D.J., Melody K., Fenton K.A., Feldmann H., Sprecher A., Zeitlin L, & Geisbert T.W. (2020). Prior vaccination with rVSV-ZEBOV does not interfere with but improves efficacy of postexposure antibody treatment. Nature Communications, 11, 3736.

Publication 2020

Thermo Autostainer 360 biotinylated goat anti-rabbit IgG

Anti antibody Anti igg Dilution Formalin Goat Hematoxylin Horseradish peroxidase Immersion Immunohistochemistry Necropsy Rabbit Streptavidin Tissues Vector

Corresponding Organization :

Other organizations : The University of Texas Medical Branch at Galveston, Office of Extramural Research, Médecins Sans Frontières

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Variable analysis

independent variables

Anti-ZEBOV VP40 primary antibody concentration (1:4000 dilution)

dependent variables

Presence and intensity of ZEBOV VP40 immunoreactivity

control variables

Tissue sample processing (immersion-fixed in 10% neutral buffered formalin and processed for histopathology)
Immunohistochemistry protocol (Thermo Autostainer 360, biotinylated goat anti-rabbit IgG secondary, Vector Horseradish Peroxidase Streptavidin, Dako DAB chromagen, Harris Hematoxylin counterstain)

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

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