Journal of General Virology

Fig. 8. Immunoblot showing incorporation of B5R protein into extracellular virions. RK₁₃ cells were infected with the indicated viruses at 10 p.f.u. per cell. At 24 h p.i. supernatants were clarified by low-speed centrifugation and then extracellular virus particles (V) were pelleted by centrifugation at 14000 r.p.m., 80 min, in a Beckman SW28.1 rotor. The supernatant (S) was removed and soluble proteins were precipitated with trichloroacetic acid and recovered by centrifugation. Cells (C) were scraped from the plastic dish into PBS and collected by centrifugation. Samples were analysed by SDS–PAGE and immunoblotting with rabbit polyclonal anti-B5R antibody. To obtain similar strength signals different amounts of samples were added for each fraction. Protein in the supernatant (S) was derived from 4x10⁵ cell equivalents, whereas extracellular virus (V) was derived from 1x10⁶ cell equivalents, and infected cell lysate was from 2x10⁴ cell equivalents. The positions of molecular mass markers in kDa are indicated.

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