Mouse monoclonal anti goat sheep igg ap gt34

Goat anti-HIV-1 gp160 (MRC ADP 72 408/5104), rabbit anti-HIV-1 p24 (Gag) (ARP 432), donkey anti-goat IgG Cy3 or FITC, and donkey anti-rabbit IgG Alexa 647 (Life Technologies, Oslo, Norway) were used for immunofluorescence assays. Goat anti-human IgG-Cy3 (Fc-specific) antibody (Sigma, Saint Louis, Missouri, USA) was used for live-cell staining and fluorescence-activated cell sorting (FACS) assay. Goat anti-HIV-1 gp120 (BioRad 5000-0557, Bio-Rad, Watford, UK), goat anti-HIV-1 p24 (Gag) (BioRad 4999–9007, Bio-Rad, Watford, UK), and mouse monoclonal anti-goat/sheep IgG–AP GT34 (Sigma, Saint Louis, Missouri, USA) were used for Western blotting. Anti-HIV-1 Env human monoclonal antibodies (MAbs) PG9, PG16, PGT128, PGT135, PGT145, CAP256-VRC26.08, VRC01, 10E8, 447-52D, and F105 were expressed in FreeStyle 293F cells (Life Technologies, Oslo, Norway) using the PEIMAX transfection reagent (Polysciences, Warrington, Pennsylvania, USA). Monoclonal antibodies were purified from cell-free supernatants after 6 days using Protein-A affinity chromatography [14 (link)]. CAP256 SU V1V2 scaffolded proteins were produced and purified by affinity chromatography using a Ni-NTA column, as described previously by Gorman et al. (2016) [15 (link)].

Goat anti-HIV-1 gp160 (MRC ADP 72 408/5104); rabbit anti-HIV-1 p24 (Gag) (ARP 432); donkey anti-goat IgG Cy3 or FITC; and donkey anti-rabbit IgG Alexa 647 (Life Technologies, Oslo, Norway) were used for immunofluorescence assays. The goat anti-human IgG-FITC (Fc-specific) antibodies (ab 97224, abcam, Cambridge, UK) were used for live-cell staining. Goat anti-HIV-1 gp120 (Bio-Rad 5000–0557, Bio-Rad, Watford, UK); goat anti-HIV-1 p24 (Gag) (Bio-Rad 4999–9007, Bio-Rad, Watford, UK); and mouse monoclonal anti-goat/sheep IgG–AP GT34 (Sigma, Saint Louis, MO, USA) were used for Western blotting. The anti-HIV-1 Env human monoclonal antibodies (MAbs) PG9, PGT128, CAP256 VRC26.08, and VRC01 were expressed in FreeStyle 293F cells (Life Technologies, Oslo, Norway) using the PEIMAX transfection reagent (Polysciences, Warrington, PA, USA). The monoclonal antibodies were purified from cell-free supernatants after 6 days using protein A affinity chromatography [39 (link)].
MBDK and HeLa cells (supplied by ATCC) were grown in Dulbecco’s Modified Eagle Medium (DMEM) (high glucose), plus L-glutamine (Lonza), plus 10% of fetal calf serum, plus 10% of penicillin-streptomycin (Pen-Strep) (both from Gibco).