Anti-RBM14 antibody [EPR29101-9]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-RBM14 antibody [EPR29101-9] (AB327067)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HepG2 cells labelling RBM14 with ab327067 at 1.0 μg/ml dilution , followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/ab150081 2μg/ml dilution (Green).

ab327067 staining RBM14 in HepG2 cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal rabbit serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab327067 at 1 μg/mL (shown in green) and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control (shown in pseudocolour magenta). Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 2 μg/mL. Nuclear DNA was labelled with DAPI (shown in blue). Also suitable in cells fixed with 100% methanol (5 min). Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/ab7291 1μg/ml 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/ab150081 2μg/ml dilution.

• WB

Lab

Western blot - Anti-RBM14 antibody [EPR29101-9] (AB327067)

Blocking and diluting buffer and concentration : 3% NFDM/TBST.

Western blot : Rabbit monoclonal [EPR29101-9] to RBM14 ab327067 staining at 1 µg/mL, shown in green; Mouse anti-GAPDH - Loading control ab8245 loading control staining at 1/20, 000 dilution, shown in magenta. A band was observed at 74 kDa. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% NFDM in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG 800CW and Goat anti-Mouse IgG 680RD at 1/20, 000 dilution.

In Western blot, Mouse anti-GAPDH - Loading control (ab8245) (1 : 20,000) (37kDa).

All lanes:

Western blot - Anti-RBM14 antibody [EPR29101-9] (ab327067) at 1 µg/mL

Lane 1:

HepG2 (human hepatocellular carcinoma epithelial cell)

Lane 2:

HEK-293 (human embryonic kidney epithelial cell)

Lane 3:

Jurkat (human T cell leukemia T lymphocyte from peripheral blood)

Lane 4:

A549 (human lung carcinoma epithelial cell)

Lane 5:

U-2 OS (human bone osteosarcoma epithelial cell)

Lane 6:

HeLa (human cervical adenocarcinoma epithelial cell)

Secondary

All lanes:

Goat Anti-Rabbit IgG H&L (800CW) and Goat Anti-Mouse IgG H&L (680RD) at 1/20000 dilution

Observed band size: 74 kDa,37 kDa

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