Anti-RhoA + C antibody [1B34A10]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RhoA + C antibody [1B34A10] (AB175359)

Immunohistochemistry analysis of Rho A/C showing staining in the cytoplasm of paraffin-embedded human hepatocarcinoma (right) compared with a negative control without primary antibody (left). To expose target proteins, antigen retrieval 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, washed with ddH2O and PBS, and then probed with ab175359 diluted in 3% BSA-PBS at a dilution of 1 : 200 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated 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

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-RhoA + C antibody [1B34A10] (AB175359)

Immunofluorescent analysis of Rho A/C (green) showing staining in the in the cytoplasm of Hela cells (right) compared to a negative control without primary antibody (left). 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 ab175359 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

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RhoA + C antibody [1B34A10] (AB175359)

mmunohistochemistry analysis of Rho A/C showing staining in the cytoplasm and membrane of paraffin-embedded human prostate carcinoma (right) compared with a negative control without primary antibody (left). To expose target proteins, antigen retrieval 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, washed with ddH2O and PBS, and then probed with ab175359 diluted in 3% BSA-PBS at a dilution of 1 : 200 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated 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

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-RhoA + C antibody [1B34A10] (AB175359)

Immunofluorescent analysis of Rho A/C (green) showing staining in the in the cytoplasm of A431 cells (right) compared to a negative control without primary antibody (left). 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 ab175359 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.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-RhoA + C antibody [1B34A10] (AB175359)

Immunofluorescent analysis of HeLa cells (formalin-fixed, 0.1% Triton X-100 permeabilized) labeling RhoA + C with ab175359 at 1/100 dilution followed with DyLight 488 goat anti-mouse IgG secondary antibody at 1/400 dilution. Nuclei (blue) were stained with Hoechst 33342 dye.

• IP

Supplier Data

Immunoprecipitation - Anti-RhoA + C antibody [1B34A10] (AB175359)

Immunoprecipitation analysis of RhoA + C was performed on HeLa cells. Antigen-antibody complexes were formed by incubating 500μg of whole cell lysate with 2μg ab175359 (lane 2).

HeLa cell lysate was run as a control (lane 1).

For WB detection, ab175359 was used at 1/1000 dilution.

All lanes:

Immunoprecipitation - Anti-RhoA + C antibody [1B34A10] (ab175359)

Predicted band size: 22 kDa

true

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RhoA + C antibody [1B34A10] (AB175359)

Immunohistochemistry analysis of Rho A/C showing staining in the cytoplasm and membrane of paraffin-embedded mouse liver tissue (right) compared with a negative control without primary antibody (left). To expose target proteins, antigen retrieval 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, washed with ddH2O and PBS, and then probed with ab175359 diluted in 3% BSA-PBS at a dilution of 1 : 100 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated 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.

• WB

Supplier Data

Western blot - Anti-RhoA + C antibody [1B34A10] (AB175359)

All lanes:

Western blot - Anti-RhoA + C antibody [1B34A10] (ab175359) at 1/1000 dilution

Lane 1:

HeLa whole cell lysate at 25 µg

Lane 2:

A431 whole cell lysate at 25 µg

Lane 3:

HEK293T whole cell lysate at 25 µg

Lane 4:

U2 OS whole cell lysate at 25 µg

Lane 5:

NIH 3T3 whole cell lysate at 25 µg

Lane 6:

K562 whole cell lysate at 25 µg

Lane 7:

RhoA purified protein at 0.5 µg

Lane 8:

RhoB purified protein at 0.5 µg

Lane 9:

RhoC purified protein with proprietary tag at 0.5 µg

Secondary

All lanes:

goat anti-mouse IgG-HRP at 1/15000 dilution

Predicted band size: 22 kDa

true

• WB

Unknown

Western blot - Anti-RhoA + C antibody [1B34A10] (AB175359)

All lanes:

Western blot - Anti-RhoA + C antibody [1B34A10] (ab175359) at 1/1000 dilution

Lane 1:

SW 480 whole cell extract at 30 µg

Lane 2:

SK-OV-3 whole cell extract at 30 µg

Lane 3:

NIH-3T3 whole cell extract at 30 µg

Lane 4:

PC-12 whole cell extract at 30 µg

Secondary

All lanes:

Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate at 1/4000 dilution

Predicted band size: 22 kDa

false