STRN3 Antibody / Striatin 3 (FY13085)

Immunofluorescent staining of STRN3 using anti-STRN3 antibody (green). STRN3 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-STRN3 antibody overnight at 4oC. DyLight 488 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 STRN3 using anti-STRN3 antibody. Lane 1: human Hela whole cell lysates, Lane 2: human MCF-7 whole cell lysates, Lane 3: human K562 whole cell lysates, Lane 4: human HepG2 whole cell lysates, Lane 5: rat brain tissue lysates, Lane 6: rat H9C2(2-1) 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-STRN3 antibody at 0.25 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. Although STRN3 isoforms are typically described at ~87 kDa and ~78 kDa, the antibody detects a predominant species at ~94 kDa across samples, with lighter lower bands. In mouse and rat brain, STRN3 appears as a doublet, consistent with differential phosphorylation within the STRIPAK scaffold that increases apparent molecular weight and resolves phospho-isoforms on SDS-PAGE. The lower signals likely represent the shorter isoform and/or limited proteolysis.

Flow Cytometry analysis of HepG2 cells using anti-STRN3 antibody. Overlay histogram showing HepG2 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-STRN3 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.