PEX11B Antibody / Peroxisomal membrane protein 11B [clone 29P66] (FY12824)

Immunohistochemical staining of PEX11B using anti-PEX11B antibody. PEX11B was detected in a paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:50 rabbit anti-PEX11B antibody overnight at 4oC. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37oC. The tissue section was developed using an HRP secondary and DAB substrate.

Western blot analysis of PEX11B using anti-PEX11B antibody. Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. Lane 1: human THP-1 whole cell lysates, Lane 2: human MCF-7 whole cell lysates, Lane 3: human Jurkat whole cell lysates, Lane 4: human REH whole cell lysates, Lane 5: human HepG2 whole cell lysates, Lane 6: mouse testis tissue lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-PEX11B antibody at 1:500 overnight at 4oC, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal was developed using an ECL Plus Western Blotting Substrate. Western blot detection of PEX11B in human cell lines reveals the expected ~25 kDa monomer band, as well as consistent higher-molecular-weight bands at ~65 kDa and ~80 kDa. The higher bands likely reflect stable PEX11B dimers/trimeric oligomers or PEX11B-containing complexes as previously documented for PEX11B self-interaction.