HLA-G Antibody (Center)

All lanes: Anti-HLA-G Antibody (Center) at 1:1000 dilution. Lane 1: Jurkat whole cell lysate. Lane 2: Raji whole cell lysate. Lane 3: Ramos whole cell lysate. Lane 4: Hela whole cell lysate. Lysates/proteins at 20 µg per lane. Secondary Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size: 38 kDa. Blocking/Dilution buffer: 5% NFDM/TBST.

Immunohistochemical analysis of paraffin-embedded Human placenta tissue was performed on the Leica BOND RXm. Tissue was fixed with formaldehyde at room temperature, antigen retrieval was by heat mediation with a EDTA buffer (pH9.0). Samples were incubated with primary antibody (1:500) for 1 hours at room temperature. A undiluted biotinylated CRF Anti-Polyvalent HRP Polymer antibody was used as the secondary antibody.

Overlay histogram showing Ramos cells stained (green line). The cells were fixed with 2% paraformaldehyde (10 min). The cells were then icubated in 2% bovine serum albumin to block non-specific protein-protein interactions followed by the antibody (1:25 dilution) for 60 min at 37°C. The secondary antibody used was Goat-Anti-Rabbit IgG, DyLight 488 Conjugated Highly Cross-Adsorbed at 1/200 dilution for 40 min at 37°C. Isotype control antibody (blue line) was rabbit IgG1 (1 μg/1x10^6 cells) used under the same conditions. Acquisition of > 10000 events was performed.