Anti-GEF H1 antibody [EPR17963] - C-terminal

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-GEF H1 antibody [EPR17963] - C-terminal (AB201687)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MCF7 (Human breast adenocarcinoma cell line) cells labeling GEF H1 with ab201687 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).

Confocal image showing cytoplasmic staining on MCF7 cell line is observed.

The nuclear counter stain is DAPI (blue).

Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows;
1. ab201687 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
2. ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-GEF H1 antibody [EPR17963] - C-terminal (AB201687)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling GEF H1 with ab201687 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).

Confocal image showing cytoplasmic staining on HeLa cell line is observed.

The nuclear counter stain is DAPI (blue).

Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows;
1. ab201687 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
2. ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-GEF H1 antibody [EPR17963] - C-terminal (AB201687)

Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed HEK293 (human embryonic kidney) cells labeling GEF H1 with ab201687 at 1/100 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and a unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.

• WB

Lab

Western blot - Anti-GEF H1 antibody [EPR17963] - C-terminal (AB201687)

Lanes 1-3 : Merged signal (red and green). Green - ab201687 observed at 112 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab201687 Anti-GEF H1 antibody [EPR17963] - C-terminal was shown to specifically react with GEF H1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265797 (knockout cell lysate ab257223) was used. Wild-type and GEF H1 knockout samples were subjected to SDS-PAGE. ab201687 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

ARHGEF knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human ARHGEF2 (GEF H1) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-arhgef2-gef-h1-knockout-hela-cell-line-ab265797'>ab265797</a>)

Lane 3:

HEK-293T cell lysate at 20 µg

Predicted band size: 111 kDa

Observed band size: 112 kDa,75 kDa

false

• WB

Supplier Data

Western blot - Anti-GEF H1 antibody [EPR17963] - C-terminal (AB201687)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687) at 1/10000 dilution

Lane 1:

HEK293 (Human embryonic kidney) lysate at 20 µg

Lane 2:

HEK293 (Human epithelial cells from cervix adenocarcinoma) lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 111 kDa

Observed band size: 112 kDa

false

Exposure time: 30s

• WB

Unknown

Western blot - Anti-GEF H1 antibody [EPR17963] - C-terminal (AB201687)

Lanes 1-3 : Merged signal (red and green). Green - ab201687 observed at 75 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab201687 Anti-GEF H1 antibody [EPR17963] - C-terminal was shown to specifically react with GEF H1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265797 (knockout cell lysate ab257223) was used. Wild-type and GEF H1 knockout samples were subjected to SDS-PAGE. ab201687 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

ARHGEF2 knockout HeLa cell lysate at 20 µg

Lane 3:

HEK293 cell lysate at 20 µg

Predicted band size: 111 kDa

Observed band size: 75 kDa

false

• WB

Supplier Data

Western blot - Anti-GEF H1 antibody [EPR17963] - C-terminal (AB201687)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687) at 1/2000 dilution

Lane 1:

Mouse brain lysate at 10 µg

Lane 2:

Rat brain lysate at 10 µg

Lane 3:

Raw264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) lysate at 10 µg

Lane 4:

C6 (Rat glial tumor cells) lysate at 10 µg

Lane 5:

NIH/3T3 (Mouse embyro fibroblast cells) at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 111 kDa

Observed band size: 112 kDa

false

Exposure time: 15s