Anti-RHOB antibody [EPR30502-160] - BSA and Azide free

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-RHOB antibody [EPR30502-160] - BSA and Azide free (AB325535)

This data was developed using ab325531, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) Daudi (human Burkitt's lymphoma lymphoblast) cells labelling RHOB with ab325531 at 1/50 (10.12 µg/ml) dilution , followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic staining in HepG2 cells and no staining in Daudi cells (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Low expression : Daudi. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-RHOB antibody [EPR30502-160] - BSA and Azide free (AB325535)

This data was developed using ab325531, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neural/glia cell cells labelling RHOB with ab325531 at 1/50 (10.12 µg/ml) dilution , followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic staining in mouse primary neural and glia cells (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection.

ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• WB

Lab

Western blot - Anti-RHOB antibody [EPR30502-160] - BSA and Azide free (AB325535)

This data was developed using ab325531, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low expression : Daudi.

Lanes 7-9 of this blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-RHOB antibody [EPR30502-160] (<a href='/en-us/products/primary-antibodies/rhob-antibody-epr30502-160-ab325531'>ab325531</a>) at 1/1000 dilution

Lane 1:

SK-BR-3 (human breast adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

SK-BR-3 transfected with siRNA specifically targeting RHOB whole cell lysate at 20 µg

Lane 3:

SK-BR-3 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

Daudi (human Burkitt's lymphoma lymphoblast) whole cell lysate at 20 µg

Lane 6:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 7:

4T1 (mouse mammary gland carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 8:

Beta-TC-6 (mouse pancreas insulinoma beta cell) whole cell lysate at 20 µg

Lane 9:

B16-F10 (mouse skin melanoma cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 23 kDa,36 kDa

true

Exposure time: 180s

• Dot

Lab

Dot Blot - Anti-RHOB antibody [EPR30502-160] - BSA and Azide free (AB325535)

This data was developed using ab325531, the same antibody clone in a different buffer formulation.

Dot blot analysis of RHOB using ab325531 at 1 : 1000 (0.506 µg/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Lane 1 : Human RHOB peptide

Lane 2 : Human RHOA peptide

Lane 3 : Human RHOC peptide

Exposure time : 180 seconds.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This antibody does not cross-react with human RHOA and RHOC .

All lanes:

Dot Blot - Anti-RHOB antibody [EPR30502-160] (<a href='/en-us/products/primary-antibodies/rhob-antibody-epr30502-160-ab325531'>ab325531</a>) at 1/1000 dilution

Lane 1:

Human RHOB peptide

Lane 2:

Human RHOA peptide

Lane 3:

Human RHOC peptide

Secondary

All lanes:

Dot Blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

false

Exposure time: 180s