Anti-Galectin 3 antibody [A3A12]

• WB

Supplier Data

Western blot - Anti-Galectin 3 antibody [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.

All lanes:

Western blot - Anti-Galectin 3 antibody [A3A12] (ab2785) at 1/1000 dilution

Lane 1:

HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 30 µg

Lane 2:

MDA-MB-231 (human breast adenocarcinoma cell line) whole cell lysate at 30 µg

Lane 3:

A549 (human lung carcinoma cell line) whole cell lysate at 30 µg

Lane 4:

PC-3 (human prostate adenocarcinoma cell line) whole cell lysate at 30 µg

Lane 5:

U-87 MG (human glioblastoma-astrocytoma epithelial cell line) whole cell lysate at 30 µg

Lane 6:

A431 (human epidermoid carcinoma cell line) whole cell lysate at 30 µg

Secondary

All lanes:

Goat anti-Mouse IgG (H+L) (HRP) at 1/4000 dilution

Predicted band size: 26 kDa

Observed band size: 30 kDa,70 kDa

true

• WB

Supplier Data

Western blot - Anti-Galectin 3 antibody [A3A12] (AB2785)

All lanes:

Western blot - Anti-Galectin 3 antibody [A3A12] (ab2785) at 1/1000 dilution

Lane 1:

MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 25 µg/mL

Lane 2:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 25 µg/mL

Lane 3:

NIH/3T3 (Mouse embryo fibroblast cell line) whole cell lysate at 25 µg/mL

Predicted band size: 26 kDa

Observed band size: 30 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Galectin 3 antibody [A3A12] (AB2785)

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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Galectin 3 antibody [A3A12] (AB2785)

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.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Galectin 3 antibody [A3A12] (AB2785)

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Galectin 3 antibody [A3A12] (AB2785)

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.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Galectin 3 antibody [A3A12] (AB2785)

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 °C. 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.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Galectin 3 antibody [A3A12] (AB2785)

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 °C. 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.

• WB

Supplier Data

Western blot - Anti-Galectin 3 antibody [A3A12] (AB2785)

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.

All lanes:

Western blot - Anti-Galectin 3 antibody [A3A12] (ab2785) at 1/1000 dilution

Lane 1:

Wild-Type HeLa (human epithelial cell line from cervix adenocarcinoma) membrane extract at 30 µg

Lane 2:

HeLa CAS9 membrane extract at 30 µg

Lane 3:

HeLa LGALS3 knockout membrane extract at 30 µg

Secondary

All lanes:

Goat anti-Mouse IgG (H+L) (HRP) at 1/4000 dilution

Predicted band size: 26 kDa

true