ARHGEF2 Antibody / Rho guanine nucleotide exchange factor 2 / GEF-H1 (FY13062)

Immunohistochemical staining of GEF-H1/ARHGEF2 using anti-ARHGEF2 antibody. GEF-H1/ARHGEF2 was detected in a paraffin-embedded section of human ovary 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-ARHGEF2 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 GEF-H1/ARHGEF2 using anti-ARHGEF2 antibody. Lane 1: human 293T whole cell lysates, Lane 2: human Hela whole cell lysates, Lane 3: human PC-3 whole cell lysates, Lane 4: rat brain tissue lysates, Lane 5: rat C6 whole cell lysates, Lane 6: mouse brain tissue lysates, Lane 7: mouse Raw264.7 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-ARHGEF2 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. A major band is observed at ~112 kDa, corresponding to full-length ARHGEF2. Additional lower-molecular-weight bands (~90-95 kDa) are commonly reported and represent truncated or dephosphorylated forms of ARHGEF2 generated by proteolytic processing or post-translational modification. These species have been described in multiple studies as part of the normal regulatory cycle of ARHGEF2.

Immunohistochemical staining of GEF-H1/ARHGEF2 using anti-ARHGEF2 antibody. GEF-H1/ARHGEF2 was detected in a paraffin-embedded section of human ovarian 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-ARHGEF2 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.

Immunohistochemical staining of GEF-H1/ARHGEF2 using anti-ARHGEF2 antibody. GEF-H1/ARHGEF2 was detected in a paraffin-embedded section of human colon 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-ARHGEF2 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.

Flow Cytometry analysis of PC-3 cells using anti-ARHGEF2 antibody. Overlay histogram showing PC-3 cells stained with (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ARHGEF2 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.