Western blot analysis of antibody responses to influenza virion proteins

Abstract

An immunoblotting procedure was developed to detect antibody responses in mice and humans to influenza virion proteins. The technique was capable of detecting 1.5 μg of haemagglutinin (HA) on nitrocellulose strips at a 1: 5000 dilution of a mouse serum with an initial haemagglutination inhibition titre of 20. The effects of the use of the blocking agent Tween‐20 on virion proteins were also studied. The commonly used concentration of 0.05% (v/v) Tween‐20, when included in blocking and incubation buffers, greatly reduced the amount of detectable matrix protein but caused no detectable loss of HA and neuraminidase/nucleoprotein proteins. If virion proteins were separated by polyacrylamide gel ..