Anti-Galectin 3抗体[A3A12] (ab2785)

Whole cell extracts were electrophoresed using a 4-12% Bis-Tris Protein Gel. Resolved proteins were then transferred onto a Nitrocellulose membrane using a dry blotting system.

Immunohistochemical analysis of human ovarian carcinoma tissue (right) labeling Galectin 3 in the nucleus and cytoplasm with ab2785, compared with a negative control (left) by Immunohistochemistry (formalin/PFA-fixed paraffin-embedded sections). To expose target proteins, antigen retrieval method was performed using 10mM sodium citrate (pH 6.0) microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature. Tissue sections were incubated with the primary antibody (1:100 in 3% BSA-PBS) overnight at 4°C. A HRP-conjugated anti-mouse IgG was used as the secondary antibody, followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Immunofluorescent analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Galectin 3 (green) in the cytoplasm and nucleus with ab2785. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Galectin 3 monoclonal antibody (Product # MA1-940) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4 °C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x

Knockout of LGALS3 was achieved by CRISPR-Cas9 genome editing using Lentiviral sgRNA and Cas9 Lentivirus. The samples were electrophoresed using 4-12% Bis-Tris Protein Gel. Resolved proteins were then transferred onto a nitrocellulose membrane using a dry blotting system.

Immunohistochemical analysis (formalin/PFA-fixed paraffin-embedded sections) human colon tissue (right) labelling Galectin 3 in the nucleus and cytoplasm with ab2785, compared with a negative control (left). To expose target proteins, antigen retrieval method was performed using 10mM sodium citrate (pH 6.0) microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature. Tissue sections were incubated with the primary antibody (1:100 in 3% BSA-PBS) overnight at 4°C. A HRP-conjugated anti-mouse IgG was used as the secondary antibody, followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Immunofluorescent analysis of NIH/3T3 (Mouse embryo fibroblast cell line) cells labelling Galectin 3 (green) in the cytoplasm and nucleus with ab2785. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were incubated with the primary antibody (1:100 in 3% BSA-PBS) overnight at 4 ºC. A DyLight-conjugated anti-mouse was used as the secondary antibody. Red (phalloidin) - F-actin, Blue - nuclei. Images were taken at a magnification of 60x.

Immunohistochemical analysis of (formalin/PFA-fixed paraffin-embedded sections) mouse colon tissue (right) labelling Galectin 3 in the nucleus and cytoplasm with ab2785, compared with a negative control (left) by . To expose target proteins, antigen retrieval method was performed using 10mM sodium citrate (pH 6.0) microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature. Tissue sections were incubated with the primary antibody (1:20 in 3% BSA-PBS) overnight at 4°C. A HRP-conjugated anti-mouse IgG was used as the secondary antibody, followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Immunofluorescent analysis of MCF7 (Human breast adenocarcinoma cell line) cells labelling Galectin 3 (green) in the cytoplasm and nucleus with ab2785. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were incubated with the primary antibody (1:100 in 3% BSA-PBS) overnight at 4 ºC. A DyLight-conjugated anti-mouse was used as the secondary antibody. Red (phalloidin) - F-actin, Blue - nuclei. Images were taken at a magnification of 60x.