LGALS3BP Antibody / Galectin-3-binding protein (FY13218)

Immunofluorescent staining of LGALS3BP using anti-LGALS3BP antibody (green). LGALS3BP was detected in an immunocytochemical section of 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-LGALS3BP 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 LGALS3BP using anti-LGALS3BP antibody. Lane 1: human Hela whole cell lysates, Lane 2: human HepG2 whole cell lysates, Lane 3: human COLO 320 whole cell lysates, Lane 4: human 293T whole cell lysates, Lane 5: rat brain 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-LGALS3BP 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. Western blot detection of LGALS3BP (Galectin-3-binding protein) shows a diffuse band pattern between ~70 and 90 kDa across multiple cell lines. Although the predicted mass is ~65 kDa, extensive N-glycosylation causes the mature secreted protein to migrate more slowly on SDS-PAGE.

Flow Cytometry analysis of HepG2 cells using anti-LGALS3BP 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-LGALS3BP 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 (Red line) was also used as a control.