Anti-Ran antibody [EPR10791(B)] - BSA and Azide free

• WB

Unknown

Western blot - Anti-Ran antibody [EPR10791(B)] - BSA and Azide free (AB249192)

We are unsure how to define the extra bands. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab155103).

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

Western blot - Anti-Ran antibody [EPR10791(B)] (<a href='/en-us/products/primary-antibodies/ran-antibody-epr10791b-ab155103'>ab155103</a>) at 1/1000 dilution

Lane 4:

Western blot - Anti-Ran antibody [EPR10791(B)] - BSA and Azide free (ab249192) 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)] - BSA and Azide free (AB249192)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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)] (<a href='/en-us/products/primary-antibodies/ran-antibody-epr10791b-ab155103'>ab155103</a>) 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

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Ran antibody [EPR10791(B)] - BSA and Azide free (AB249192)

Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Ran with purified ab155103 at 1/100 dilution (10 μg/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).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab155103).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Ran antibody [EPR10791(B)] - BSA and Azide free (AB249192)

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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab155103).

• IP

Unknown

Immunoprecipitation - Anti-Ran antibody [EPR10791(B)] - BSA and Azide free (AB249192)

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
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab155103).

All lanes:

Immunoprecipitation - Anti-Ran antibody [EPR10791(B)] (<a href='/en-us/products/primary-antibodies/ran-antibody-epr10791b-ab155103'>ab155103</a>)

Predicted band size: 24 kDa,43 kDa

false