All lanes : Anti-eEF1A1/EF-Tu antibody [EPR9470] (ab140632) at 1/1000 dilution

Lane 1 : HeLa cell lysate
Lane 2 : MCF-7 cell lysate
Lane 3 : 293T cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP labelled
Goat anti-Rabbit at 1/2000 dilution

Predicted band size: 50 kDa

Immunocytochemistry/Immunofluorescence analysis of MCF-7 cells labelling eEF1A1/EF-Tu with purified ab140632 at a dilution of 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Nuclei counterstained with DAPI (blue).

Control: PBS only.

Immunohistochemical analysis of paraffin embedded Human brain tissue labelling eEF1A1/EF-Tu with ab140632 at 1/50 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Immunohistochemical analysis of paraffin embedded Human breast carcinoma tissue labelling eEF1A1/EF-Tu with ab140632 at 1/50 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Overlay histogram showing MCF7 cells stained with ab140632 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab140632, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor^® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in MCF7 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.