VOPBAVirus Overlay Protein Binding Assay
VOPBAVirus Overlay Protein Blot Assay
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In VOPBA, the protein interactions with the virus particles occurred under the reduced conditions that are required for the SDS-PAGE; therefore, to detect native proteins in CNS gray matter lysates interacting with E protein, we used coimmunoprecipitation assays.
Additionally, a 130 kDa membrane protein of human hepatocytes (HEpG2 cell line) was shown to bind DENV particles of serotypes 3 and 4, but not 2 by VOPBA [55].
For the VOPBA, the membrane proteins were processed (reduced with fi-mercaptoethanol) and electrophoresed in SDS-PAGE; thus no native proteins were involved in the interaction with the virus particle.
Marissa Perez-Garcia established the primary-CNS cultures and performed the VOPBA and immunofluorescence assays.