Journal:

Article Title: Recombinant Rinderpest Vaccines Expressing Membrane-Anchored Proteins as Genetic Markers: Evidence of Exclusion of Marker Protein from the Virus Envelope

doi:

Figure Lengend Snippet: Analysis of HA and ANC-GFP expression and secretion by RPVINS-HA- and RPVANC-GFP-infected B95a cells. Cells were labeled for 2 h and then the medium was replaced with normal medium containing unlabeled methionine and cysteine. Medium was collected after 0, 1, 2, 4, 6, and 8 h of chase and analyzed for the presence of labeled secreted proteins, whereas the corresponding cell sheet was lyzed and analyzed for the presence of labeled intracellular proteins. Proteins were immunoprecipitated with either anti-HA polyclonal antibody (αHA), anti-GFP polyclonal antibody (αGFP), or anti-RPV P monoclonal antibody (αRPV-P) as described in Materials and Methods. (A to D) RPVINS-HA-infected cells; (E to H) RPVANC-GFP-infected cells. Numbers above the gels represent the time in hours after protein labeling at which the samples were collected. Positions of the molecular mass markers in kilodaltons are indicated on the left. Positions of the RPV N and P proteins are indicated on the right.

Article Snippet: Antibodies used were 0.5 μl of rabbit anti-HA polyclonal antibody (provided by D. A. Steinhauer), 0.5 μl of polyclonal rabbit anti-GFP antibody (Clontech), and 0.2 μl of mouse anti-RPV P (phosphoprotein) monoclonal antibody 2-1 ( 5 ) together with 0.5 μl of rabbit anti-mouse antibody (Dakopatts).

Techniques: Expressing, Infection, Labeling, Immunoprecipitation