Flow Cytometry analysis of in human peripheral blood mononuclear cells labelling T-Bet/TBX21 with ab275959 (right) or isotype control (left). Cells were fixed with 4% paraformaldehyde (10 min RT, 1 hr on ice) and stored in 80% methanol overnight (-20°C). The cells were then permeabilized with 0.2% Tween 20 in PBS containing 0.5% BSA. 0.2µg of ab275959 was added per 1x10E6 cells and incubated for 30 min on ice. DyLight® 488 goat anti-rabbit IgG secondary antibody at 1/1000 dilution was used as the secondary antibody. Isotype control (left) was purified rabbit IgG.

Flow Cytometry analysis of in MJ[G11] cells labelling T-Bet/TBX21 with ab275959 (red shaded) or isotype control (unshaded). DyLight® 488-conjugated goat anti-rabbit IgG was used as a secondary antibody.

All lanes : Anti-T-bet / Tbx21 antibody [BLR110H] (ab275959) at 1/1000 dilution

Lane 1 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
Lane 2 : HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
Lane 3 : MJ/G11 (Human T lymphocyte cell line) whole cell lysate
Lane 4 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 5 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 6 : NK-92 (Human peripheral blood malignant non-Hodgkin's lymphoma cell line) whole cell lysate
Lane 7 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
Lane 8 : U-2 OS (Human bone osteosarcoma epithelial cell line) whole cell lysate

Lysates/proteins at 50 µg per lane.

Secondary
All lanes : HRP conjugated goat anti-rabbit IgG

Predicted band size: 58 kDa

Chemiluminescence with an exposure time of 30 seconds.

Lower Panel: Rabbit anti-Actin recombinant monoclonal.

Paraffin embedded Hodgkin's lymphoma tissue stained for T-bet / Tbx21 using ab275959 at 1/500 dilution in immunohistochemical analysis. Secondary antibody HRP-conjugated goat anti-rabbit IgG was used. DAB substrate.

MJ (Primary cutaneous T-cell non-Hodgkin lymphoma) cells stained for T-bet / Tbx21 using ab275959 at 1/500 dilution in ICC. Secondary antibody HRP-conjugated goat anti-rabbit IgG was used. DAB substrate.

Whole cell lysate (1.0 mg per IP reaction; 20% of IP loaded) from MJ/G11 cells
prepared using NETN lysis buffer. ab275959 was used for immunoprecipitation at 6 µl per reaction. For blotting ab275959 was used at 1/1000 dilution.

Lane 1: ab275959.

Lane 2: Control IgG.

Chemiluminescence with an exposure time of 3 seconds.

Paraffin embedded seminoma tissue stained for T-bet / Tbx21 using ab275959 at 1/500 dilution in immunohistochemical analysis. Secondary antibody HRP-conjugated goat anti-rabbit IgG was used. DAB substrate.