Anti ha monoclonal antibody clone ha 7

The primary antibodies used were as follows: mouse monoclonal anti-HSV gB (Virusys), mouse monoclonal anti-HSV1 gC clone 3G9 (Abcam), rabbit polyclonal anti-LC3B (Cell Signaling Technology), mouse monoclonal anti-β-actin, anti-FLAG M2 monoclonal, and anti-HA monoclonal antibody, clone HA-7 (Sigma-Aldrich). For Western blotting, the secondary antibodies used were peroxidase-conjugated AffiniPure F(ab′)2 fragment of goat anti-mouse IgG(H + L) or peroxidase-conjugated AffiniPure F(ab′)2 fragment of goat anti-rabbit IgG(H + L) (Jackson ImmunoResearch). Secondary antibodies for immunofluorescence were AlexaFluor 594 F(ab′)2 fragment of goat anti-mouse IgG(H + L) or AlexaFluor 488 F(ab′)2 fragment of goat anti-rabbit IgG(H + L) (Life Technologies).

The following gB-specific antigens, derived from HCMV strain AD169, were used: AD1 containing aa 484–650, was expressed with galactosidase as a fusion partner in Escherichia coli. The construction of galactosidase-containing plasmids has been described in detail elsewhere [30 (link)].
AD2, a short linear peptide containing aa 68–80, was synthesized chemically, as described in detail elsewhere [30 (link), 33 (link)].
AD4 contained a fused polypeptide of aa 121–132 and 344–438. For determination of AD4-specific antibodies a purified GST–AD4 fusion protein was used as antigen and expressed in E. coli, as described by Spindler et al [35 (link)].
AD5 contained aa 133 to 343. AD5-specific antibodies were determined in a capture enzyme-linked immunosorbent assay (ELISA) using a mammalian cell (HEK 293T) derived AD5 polypeptide containing an HA epitope tag at the amino terminus of the protein, as described elsewhere [36 (link)]. To capture the antigen, an anti-HA monoclonal antibody (clone HA-7, Sigma-Aldrich) was diluted to 1 µg/mL in 0.05 M sodium carbonate buffer pH 9.6, and 50 µL/well was used to coat polystyrene 96-well plates (NuncImmuno) overnight at 4°C.