Anti-Ran antibody [EPR10791(B)]

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Ran antibody [EPR10791(B)] (AB155103)

Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Ran with purified ab155103 at 1/100 dilution (10μg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Ran antibody [EPR10791(B)] (AB155103)

Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Ran with purified ab155103 at 1/100 dilution (10 μg/mL). Cells were fixed in 100% Methanol and permeabilized with None. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/1000 (2 μg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• IP

Unknown

Immunoprecipitation - Anti-Ran antibody [EPR10791(B)] (AB155103)

Purified ab155103 at 1/50 dilution (2 μg) immunoprecipitating Ran in HeLa whole cell lysate.
Lane 1 (input) : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10 μg
Lane 2 (+) : ab155103 + HeLa whole cell lysate.
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab155103 in HeLa whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration : 5% NFDM/TBST.
Diluting buffer and concentration : 5% NFDM/TBST.
Observed band size : 24 kDa

All lanes:

Immunoprecipitation - Anti-Ran antibody [EPR10791(B)] (ab155103)

Predicted band size: 24 kDa,43 kDa

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• WB

Unknown

Western blot - Anti-Ran antibody [EPR10791(B)] (AB155103)

We are unsure how to define the extra bands.

Lanes 1, 2, 3, 5, 6, 7 and 8:

Western blot - Anti-Ran antibody [EPR10791(B)] (ab155103) at 1/1000 dilution

Lane 4:

Western blot - Anti-Ran antibody [EPR10791(B)] - BSA and Azide free (<a href='/en-us/products/primary-antibodies/ran-antibody-epr10791b-bsa-and-azide-free-ab249192'>ab249192</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

LoVo (Human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate at 20 µg

Lane 4:

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

Lane 5:

C6 (Rat glial tumor glial cell) whole cell lysate at 20 µg

Lane 6:

Raw 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 7:

PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg

Lane 8:

NIH/3T3 (Mouse embryonic fibroblast) 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

Predicted band size: 24 kDa

Observed band size: 24 kDa

false

• WB

Lab

Western blot - Anti-Ran antibody [EPR10791(B)] (AB155103)

Blocking/Diluting buffer and concentration 5% NFDM/TBST The bands observed at 30kDa, 60kDa and 150kDa should be monomer, dimer and polymer respectively.

All lanes:

Western blot - Anti-Ran antibody [EPR10791(B)] (ab155103) at 1/1000 dilution

Lane 1:

HEK-293T transfected with an empty vector whole cell lysate at 20 µg

Lane 2:

HEK-293T transfected with a human Ran expression vector containing a Flag tag 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: 24 kDa,30 kDa,60 kDa,150 kDa

false

Exposure time: 7s