Anti-Firefly Luciferase antibody [EPR17790]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Firefly Luciferase antibody [EPR17790] (AB185924)

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 Firefly Luciferase, labeling Firefly Luciferase with ab185924 at 1/500 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 Firefly 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;
-ve control 1 : ab185924 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 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 Cyt

Lab

Flow Cytometry - Anti-Firefly Luciferase antibody [EPR17790] (AB185924)

Flow Cytometry analysis of 293T (Human epithelial cell line from embryonic kidney) cells transfected with GFP tagged Firefly Luciferase labeled with purified ab185924 at 1/70 dilution (10μg/ml, Right panel). 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.

• WB

Supplier Data

Western blot - Anti-Firefly Luciferase antibody [EPR17790] (AB185924)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Firefly Luciferase antibody [EPR17790] (ab185924) at 1/1000 dilution

Lane 1:

Firefly Luciferase transfected HEK293 (human embryonic kidney) whole cell lysate at 10 µg

Lane 2:

HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 3:

MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 4:

Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate at 20 µg

Lane 5:

SH-SY5Y (Human neuroblastoma from bone marrow cells) whole cell lysate at 20 µg

Lane 6:

Human fetal brain lysate at 10 µg

Lane 7:

Human fetal heart lysate at 10 µg

Lane 8:

Human fetal kidney lysate at 10 µg

Lane 9:

Human fetal spleen lysate at 10 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 61 kDa

Observed band size: 61 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-Firefly Luciferase antibody [EPR17790] (AB185924)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Firefly Luciferase antibody [EPR17790] (ab185924) at 1/1000 dilution

Lane 1:

Firefly Luciferase transfected HEK293 (Human embryonic kidney) whole cell lysate at 10 µg

Lane 2:

Mouse brain lysate at 10 µg

Lane 3:

Mouse heart lysate at 10 µg

Lane 4:

Mouse kidney lysate at 10 µg

Lane 5:

Mouse spleen lysate at 10 µg

Lane 6:

Rat brain lysate at 10 µg

Lane 7:

Rat heart lysate at 10 µg

Lane 8:

Rat kidney lysate at 10 µg

Lane 9:

Rat spleen lysate at 10 µg

Lane 10:

C6 (Rat glial tumor cells) whole cell lysate at 10 µg

Lane 11:

RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg

Lane 12:

PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate at 10 µg

Lane 13:

NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution

Predicted band size: 61 kDa

Observed band size: 61 kDa

false

Exposure time: 3min

• WB

Unknown

Western blot - Anti-Firefly Luciferase antibody [EPR17790] (AB185924)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Firefly Luciferase antibody [EPR17790] (ab185924) at 1/20000 dilution

Lane 1:

Empty vector (vector control) transfected HEK293 (Human embryonic kidney) whole cell lysate at 10 µg

Lane 2:

Firefly Luciferase transfected HEK293 (Human embryonic kidney) whole cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution

Predicted band size: 61 kDa

Observed band size: 61 kDa

false

Exposure time: 3min