STOML2 Antibody / Stomatin-like protein 2 (FY12028)

IHC staining of STOML2 using anti-STOML2 antibody. STOML2 was detected in a paraffin-embedded section of human colon 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 2 ug/ml rabbit anti-STOML2 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 STOML2 using anti-STOML2 antibody. Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. Lane 1: human 293T whole cell lysates, Lane 2: human Hela whole cell lysates, Lane 3: human Jurkat whole cell lysates, Lane 4: human MCF-7 whole cell lysates, Lane 5: rat brain tissue lysates, Lane 6: rat C6 whole cell lysates, Lane 7: mouse brain tissue lysates, Lane 8: mouse NIH/3T3 whole cell 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-STOML2 antibody at 0.5 ug/ml 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. The expected band size for STOML2 is at 39 kDa.

Immunoprecipitating STOML2 in Jurkat whole cell lysate. Western blot analysis of STOML2 using anti-STOML2 antibody. Lane 1: jurkat whole cell lysates (30ug), Lane 2: Rabbit control IgG instead of anti-STOML2 antibody in jurkat whole cell lysate, Lane 3: anti-STOML2 antibody (2ug) + jurkat whole cell lysate (500ug). After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-STOML2 antibody at a dilution of 0.5 ug/ml and probed with a goat anti-rabbit IgG-HRP secondary antibody. The signal is developed using ECL Plus Western Blotting Substrate. The expected band size for STOML2 is at 39 kDa.

Flow Cytometry analysis of Jurkat cells using anti-STOML2 antibody. Overlay histogram showing Jurkat cells stained with (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-STOML2 antibody (1 ug/million cells) for 30 min at 20oC. DyLight 488 conjugated goat anti-rabbit IgG (5-10 ug/million cells) was used as secondary antibody for 30 minutes at 20oC. Isotype control antibody (Green line) was rabbit IgG (1 ug/million cells) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.