STING1 Antibody Rabbit Polyclonal for IF / STING1 Immunofluorescence Antibody (FY13022)

Immunohistochemical staining of TMEM173/STING using anti-STING1 antibody. TMEM173/STING was detected in a paraffin-embedded section of human lung 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-STING1 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 TMEM173/STING using anti-STING1 antibody. Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. Lane 1: human Jurkat whole cell lysates, Lane 2: mouse spleen tissue lysates, Lane 3: mouse thymus tissue lysates, Lane 4: mouse MH-S 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-STING1 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. A prominent band is detected at approximately 37 kDa, slightly below the predicted molecular weight of 42 kDa. This migration pattern is consistent with published reports showing that the transmembrane nature of STING1 leads to faster electrophoretic mobility on SDS-PAGE.

STING1 Antibody Rabbit Polyclonal for IF immunofluorescence analysis of human cells. Immunofluorescent staining of human SiHa cells using STING1 Antibody Rabbit Polyclonal for IF demonstrates cytoplasmic fluorescence consistent with intracellular localization of Stimulator of interferon genes protein / STING1 (TMEM173). STING1 signal is visualized in green using DyLight 488-conjugated secondary antibody, while Beta Tubulin staining appears red following detection with Cy3-conjugated secondary antibody, highlighting the cytoskeletal network. Cells were blocked with 10% goat serum and incubated with rabbit anti-STING1 antibody (5 ug/ml) together with mouse anti-Beta Tubulin antibody overnight at 4oC. Following secondary antibody incubation, fluorescence signals were visualized by microscopy using filter sets appropriate for the fluorescent labels.

Flow Cytometry analysis of HepG2 cells using anti-STING1 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-STING1 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.