STIM2 Antibody / Stromal interaction molecule 2 (FY13386)

Immunofluorescent staining of STIM2 using anti-STIM2 antibody (red). STIM2 was detected in an immunocytochemical section of human HeLa cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 ug/ml rabbit anti-STIM2 antibody overnight at 4oC. Cy3 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37oC. The section was counterstained with DAPI nuclear stain (blue). Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Western blot analysis of STIM2 using anti-STIM2 antibody. Lane 1: human Hela whole cell lysates, Lane 2: human 293T whole cell lysates, Lane 3: human Jurkat whole cell lysates, Lane 4: human HepG2 whole cell lysates, Lane 5: rat PC-12 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-STIM2 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 enhanced chemiluminescent. STIM2 antibody detects a predominant band at approximately 100 kDa in human HeLa, 293T, Jurkat, and HepG2 lysates, slightly above the 84 kDa theoretical mass but consistent with the 100-115 kDa phosphorylated and glycosylated STIM2 isoforms described in the literature. Rat PC-12 lysate shows a modestly faster migrating band within the same range, in line with species- and phosphorylation-dependent mobility differences.