All lanes : Anti-Renilla Luciferase antibody [EPR17792] (ab185926) at 1/1000 dilution (Purified)

Lane 1 : 293T (Human embryonic kidney epithelial cell) transfected with an empty vector (vector control), whole cell lysate
Lane 2 : 293T (Human embryonic kidney epithelial cell) transfected with Renilla Luciferase expression vector, whole cell lysate

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 36 kDa
Observed band size: 36 kDa

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK293T (Human epithelial cells from embryonic kidney) cells transfected with Empty vector or Renilla Luciferase, labeling Renilla Luciferase with ab185926 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Nuclear staining on HEK293T cells transfected with Renilla Luciferase 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. ab185926 at 1/1000 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 Cytometry analysis of 293T (Human epithelial cell line from embryonic kidney) transfected with GFP tagged Renilla Luciferase cells labeled with purified ab185926 at 1/70 dilution (10µg/ml, Right panel) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 647)(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Left panel) was used as the isotype control.