Journal:
Article Title: Chimeric Nectin1-Poliovirus Receptor Molecules Identify a Nectin1 Region Functional in Herpes Simplex Virus Entry
doi: 10.1128/JVI.75.17.7987-7994.2001
Figure Lengend Snippet: FACS analysis of immunoreactivity to R1.302 and anti-PVR MAbs and of gD binding. Cells were incubated either with R1.302 or PV.404 MAbs or with gD(Δ290-299t) followed by anti-gD MAb H170. Except for nectin1α and -β, all cultures were subjected to G418 selection followed by FACS sorting. After FACS sorting and culturing, the cells exhibited heterogeneous fluorescence profiles. This most likely reflects cell populations with heterogeneity in the extent of expression. Cell surface expression analysis: dashed line, negative control for MAb reactivity consisting of isotype-matched irrelevant MAb, followed by secondary antibody; thin line, anti-PVR MAb PV.404; thick line, anti-Nectin1 MAb R1.302. gD-binding analysis: dashed line, negative control consisting of no gD, followed by MAb H170; thick line, gD at 0.5 μg/ml.
Article Snippet: For FACS analysis of immunoreactivity to R1.302 and anti-PVR antibodies and analysis of gD binding, live cells were incubated with the following antibodies: P44 and P242 hybridoma cell supernatants at 1:2 dilution, MAb 280 ascitic fluid at 1:1,000 dilution, purified D171 and PV.404 at 10 μg/ml, or 0.5 μg of recombinant gD(Δ290-299t)/ml and mouse anti-gD MAb H170 (Goodwin Cancer Research Institute, Plantation, Fla.) (1:1,000), followed by a 1:50 goat anti-mouse phycoerythrin antibody (Beckman-Coulter-Immunotech, France).
Techniques: Binding Assay, Incubation, Selection, Fluorescence, Expressing, Negative Control